Treatment of atherosclerosis with cholesterol ester transport protein mimotopes

ABSTRACT

The present invention relates to the use of compounds for producing a medicament for preventing and/or treating atherosclerosis, atherosclerosis risk diseases and atherosclerosis sequelae.

This application is a divisional of U.S. Ser. No. 12/673,081 filed Feb.11, 2010, incorporated herein by reference, which was a National Stageof PCT/AT08/000,281 filed Aug. 8, 2008 and claims the benefit ofAustrian application A1 258/2007 filed Aug. 10, 2007.

The invention relates to the prevention and treatment ofatherosclerosis, atherosclerosis risk diseases and atherosclerosissequelae.

Atherosclerotic sequelae, such as the peripheral arterial occlusiondisease, coronary heart disease as well as the apoplectic cerebralinsultus, are still among the main causes of death in the United States,Europe, and in large parts of Asia. The development of theatherosclerosis is considered to be a chronic progressive inflammationof the arterial vessel wall which is characterized by a complexinteraction of growth factors, cytokines and cell interactions.According to the “response-to-injury” hypothesis, the “injury” of theendothelium constitutes the initial event of the disease, leading to anendothelial dysfunction which triggers a cascade of cellularinteractions culminating in the formation of the atheroscleroticlesions. As risk factors promoting such an “injury”, exogenous andendogenous influences are mentioned which correlate statisticallysignificantly with atherosclerosis. Increased and modified LDL, Lp(a),arterial hypertension, Diabetes mellitus and hyperhomocysteinaemia are,for instance, counted among the most important ones of theseendothelium-damaging factors. Since the endothelium does not constitutea rigid, but much rather an extremely dynamic barrier, a plurality ofmolecular changes occur in the course of the endothelial dysfunction inaddition to an increased permeability for lipoproteins, which molecularchanges have a decisive influence on the interaction of monocytes,T-lymphocytes and endothelial cells. By the expression of endothelialadhesion molecules of the type of the E, L and P selectins, integrins,ICMA-1, VCAM-1 and platelet-endothelial-cell adhesion molecule-1,adhesion of monocytes and T-lymphocytes at the lumen side occurs. Thesubsequent migration of the leukocytes over the endothelium is mediatedby MCP-1, interleukin-8, PDGF, MCSF and osteopontin. Via the so-calledscavenger receptor, macrophages and monocytes resident in the intima arecapable of taking up the penetrated LDL particles and to deposit them asvacuoles of cholesterol esters in the cytoplasma. The foam cells formedin this manner accumulate mainly in groups in the region of the vesselintima and form the “fatty streak” lesions occurring already inchildhood. LDL are lipoproteins of low density and are formed bycatabolic effects of lipolytic enzymes from VLDL particles rich intriglyceride. Besides their damaging properties on endothelial cells andsmooth muscle cells of the media, LDL moreover has a chemotactic effecton monocytes and is capable of increasing the expression of MCSF andMCP-1 of the endothelial cells via gene amplification. In contrast toLDL, HDL is capable of taking up cholesterol esters from loadedmacrophages mediated by apolipoprotein E, under formation of so-calledHDLc complexes. By the interaction of SR-B1 receptors, these cholesterolester-loaded particles are capable of binding to hepatocytes or to cellsof the adrenal cortex and delivering cholesterol for the production ofbile acids and steroids, respectively. This mechanism is called reversecholesterol transport and elucidates the protective function of HDL.Activated macrophages are capable of presenting antigens via HLA-DR andthereby activate CD4 and CD8 lymphocytes which, consequently, arestimulated to secrete cytokines, such as IFN-gamma and TNF-alpha, andmoreover, contribute to increasing the inflammatory reaction. In thefurther course of the disease, smooth muscle cells of the media start togrow into the region of the intima which has been altered byinflammation. By this, the intermediary lesion forms at this stage.Starting from the intermediary lesion, the progressive and complicatedlesion will develop over time, which is morphologically characterized bya necrotic core, cellular detritus and a fibrinous cap rich in collagenon the side of the lumen. If the cell number and the portion of thelipoids increase continuously, tears in the endothelium will occur, andsurfaces with thrombotic properties will be exposed. Due to the adhesionand activation of thrombocytes at these tears, granules will be releasedwhich contain cytokines, growth factors and thrombin. Proteolyticenzymes of the macrophages are responsible for the thinning of thefibrinous cap which, at last, will lead to a rupture of the plaques withconsecutive thrombosis and stenosing of the vessels and an acuteischemia of the terminal vessels.

Various risk factors are held responsible for the forming ofatherosclerotic lesions. Hyperlipoproteinemia, arterial hypertension andabuse of nicotine are of particular significance in this respect. Adisease which involves an excessive increase in the total and LDLcholesterol is the familial hypercholesterinemia (FH). It belongs to themost frequent monogenetically inherited metabolic diseases. The moderateheterozygous form occurs with a frequency of 1:500, the homozygous formwith 1:1 million clearly more rarely. Causes of the familialhypercholesterinemia are mutations in the LDL receptor gene on the shortarm of chromosome 19. These mutations may be deletions, insertions orpoint mutations. The characteristic finding of the lipoproteins infamilial hypercholesterinemia is an increase in the total and LDLcholesterol at mostly normal triglyceride and VLDL concentrations. Oftenthe HDL is lowered. Phenotypically, there is a typeIIAa-hyperlipoproteinemia. In the heterozygous form, the totalcholesterol is increased by the two to three-fold, in the homozygousform it is increased by the five to six-fold as compared to the normallevel. Clinically the familial hypercholesterinemia manifests itself byan early coronary sclerosis. As a rule, in heterozygous men the firstsymptoms of a coronary heart disease (CHD) occur between their 30^(th)and the 40^(th) year of age, in women on an average 10 years later. 50%of the afflicted men die of the consequences of their coronary sclerosisbefore they are 50 years old. Besides the massively increased LDLlevels, also lowered HDL concentrations are responsible for the rapidprogress of atherosclerosis. Atherosclerotic changes may become manifestalso on extra-cardiac vessels, such as the aorta, the carotid arteriesand peripheral arteries. With the homozygous form of the disease, thecoronary sclerosis develops already in early childhood. The firstmyocardial infarction often occurs before the 10^(th) year of age, andin most cases the afflicted persons die before they are 20 years old.The development of xanthomas is a function of the level of the serumcholesterol and the duration of the disease. Approximately 75% of theheterozygous individuals afflicted who are more than 20 years oldexhibit tendinous xanthomas. The homozygous individuals have skin andtendon xanthomas in nearly 100%. Lipid deposits may also occur on theeye lid and in the cornea (xanthelasmas; Arcus lipoides). These are,however, not a specific sign of a hypercholesterinemia, since they arealso found with normal cholesterol levels. Furthermore, with the FH,acute arthritides and tendosynovitides occur frequently. The individuallipoproteins differ with respect to size and density, since they containdifferently large portions of lipids and proteins, so-calledapoproteins. The density increases with increasing protein anddecreasing lipid portion. Due to their different densities, they can beseparated into different fractions by ultracentrifugation. This is thebasis for the classification of the lipoproteins into their main groups:chylomicrones, very-low-density lipoproteins (VLDL),intermediate-density lipoproteins (IDL), low-density lipoproteins (LDL),high-density lipoproteins (HDL), lipoprotein (a) (Lp(a)). Among thelipoproteins with a high atherogenic potential there are primarily theLDL, the Lp(a) and the VLDL. LDL has a density of approximatelyd=1.006-1.063 g/ml. The core is formed by esterified cholesterolmolecules. This highly hydrophobic core is surrounded by an envelope ofphospholipids, non-esterified cholesterol and one single Apo B100molecule. Besides, Apoprotein E is found on the surface of the LDLparticles. The function of the LDL consists in transporting cholesterolto peripheral tissues where—mediated by the apoprotein B-100—it is takenup into the cells via the LDL receptor. In comprehensive epidemiologicstudies, a positive correlation between the level of the serumcholesterol and the occurrence of a coronary heart disease could bedemonstrated. LDL cholesterol levels of higher than 160 mg/dl constitutea high cardiovascular risk. Besides the level of the LDL cholesterol,also the level of the vessel-protecting HDL cholesterol plays animportant role when estimating the risk profile for cardiovasculardiseases. Levels of below 35 mg/dl are associated with an increasedrisk. VLDL are lipoproteins with a low density (d=0.94-1.006 g/ml) and ahigh triglyceride portion. Substantially, VLDL contain apoprotein C, andsmall portions of apoproteins B-100 and E. Different from chylomicrons,VLDL do not consist of food lipids, but are synthesized in the liverfrom endogenously formed triglycerides and secreted into circulation. Aswith the chylomicrons, the triglycerides are hydrolyzed by theaproprotein C-II-activated lipoprotein-lipase, and the free fatty acidsare supplied to the muscle and fat tissue. The remainingcholesterol-rich VLDL remnants are called intermediate densitylipoproteins because of their higher density. Lipoprotein(a) (Lp(a)) hasa density of 1.05 to 1.12 g/ml and resembles LDL in its composition.Besides apoprotein B-100, its protein portion consists of theapoprotein(a) which is characteristic of Lp(a). To date, very little isknown about the physiology and function of the Lp(a). Since theapoprotein(a) molecule has a high sequence homology to plasminogen, itis assumed that Lp(a) both promotes the formation of thrombi onatherosclerotic plaques and also has an atherogenic effect. Lp(a) isfound together with apoprotein B in atherosclerotic lesions.Retrospective studies have shown a correlation between increased Lp(a)and a CHD. Likewise, the metaanalysis of numerous prospective studieshas shown that Lp(a) is an independent risk factor for the occurrence ofa CHD. Levels of between 15 and 35 mg/dl are considered to be normal. Sofar, Lp(a) can be influenced neither by diet nor by medicaments.Therefore, therapy measures are restricted to reducing further riskfactors. In particular, a lowering of the LDL cholesterol seems to lowerthe cardiovascular risk of Lp(a). In the pathogenesis ofatherosclerosis, considerable pathophysiologic importance is, moreover,attributed to coagulation factors. Epidemiologic findings suggest acorrelation between the fibrinogen concentration in plasma and thedevelopment of a coronary heart disease, and, primarily, a myocardialinfarction. In this context, increased fibrinogen levels (>300 mg/dl)proved to be an independent indicator and risk factor for cardiovasculardiseases. Yet also high concentrations of the tissue plasminogenactivator inhibitor tPA-I are associated with the occurrence of CHD. Therelationship between hypertriglyceridemia and coronary risk is adifferent one in each case, depending on the cause of the elevation ofthe blood lipids. Despite the discussion whether or not triglyceridesare to be considered as an independent risk factor it is undisputed thatthey play an important role in the pathogenesis of coronary heartdiseases. Incidence of the disease is the highest in patients whoexhibit high LDL cholesterol and a high triglyceride level.

The cholesterol ester transfer protein (CETP) is a stable plasmaglycoprotein which is responsible for the transfer of neutral lipids andphospholipids between lipoproteins and which down-regulates the plasmaconcentration of HDL. The inhibition of the CETP lipid transfer activityhas already been suggested as a therapeutic approach for increasing theHDL plasma level. There are numerous reasons which suggest that thereduction of CETP activity in plasma should lead to an increase in theHDL levels. Thus, CETP lowers the HDL concentration by the transfer ofcholesterol esters from HDL to LDL and VLDL. In animal experiments withrabbits and hamsters, the transient inhibition of CETP with anti-CETPmonoclonal antibodies, antisense oligonucleotides or CETP inhibitors ledto the increase in the HDL levels. Lasting CETP inhibition withantisense oligonucleotides increased the HDL levels and, thus, led to areduction of the atherosclerotic lesions in the rabbit animal model foratherosclerosis.

In the literature several CETP inhibitors are described, some of whichare in clinical trials (e.g. Anacetrapib (Krishna R., Lancet 370 (9603)(2007): 1907-14) and Torcetrapib (Sikorski, J. A., J. Med. Chem. 49 (1)(2006): 1-22)).

In U.S. Pat. No. 5,512,548 and in WO 93/011782, polypeptides and theiranalogues are described which are capable of inhibiting CETP thatcatalyses the transfer of cholesterol esters from HDL to VLDL and LDL,and, therefore, have anti-atherosclerotic activity if administered to apatient. According to these documents, such a CETP polypeptide inhibitoris derived from apolipoprotein C-I of various sources, whereinespecially N-terminal fragments up to amino acid 36 have been identifiedas CETP inhibitors.

Also in U.S. Pat. No. 5,880,095 A, a CETP-binding peptide is disclosedwhich is capable of inhibiting the activity of CETP in an individual.The CETP-inhibitory protein comprises an N-terminal fragment of porcineapolipoprotein C-III.

In the US 2006/0276400 and the WO 96/034888 peptides are disclosed,which are derived from CETP and comprise T-cell and/or B-cell epitopes.These peptides are able to induce in vivo the formation of CETP specificantibodies.

In US 2004/0087481 and U.S. Pat. No. 6,410,022 B1, peptides aredisclosed which, because of the induction of a CETP-specific immuneresponse, can be used for the treatment and prevention of cardiovasculardiseases, such as, e.g., atheroslerosis. These peptides comprise a Thelper cell epitope which is not derived from CETP, and at least oneB-cell epitope that comes from CETP and can be derived directly from thelatter. The T helper cell epitope advantageously is derived from tetanustoxoid and is covalently bound to at least one B-cell epitope of CETP.By using a T helper cell epitope that is alien to the organism, itbecomes possible to induce antibodies in the body of an individual,which antibodies are directed against that peptide portion that consistsof at least one CETP-B-cell epitope.

In Mao D et al (Vaccine 24 (2006): 4942-4950) the use of a plasmidcomprising a nucleic acid molecule encoding for a B cell epitope of CETPas vaccine is described.

In the WO 2006/029982 CETP mimotopes to be used for the manufacture of amedicament for the treatment or prevention of atherosclerosis isdescribed.

Most recently, there have already been suggestions for a vaccineapproach with regard to CETP. Thus, e.g., rabbits have been treated witha vaccine which contained that peptide of CETP responsible for thecholesterol-ester transfer as an antigen. The immunized rabbits had areduced CETP activity and altered lipoprotein levels with increased HDLand reduced LDL values. Moreover, the treated test animals of theatherosclerosis model also showed reduced atherosclerotic lesions incomparison with control animals.

The results of a phase II-clinical study were published, which study hadbeen carried out by the American biotechnology company Avant with thevaccine CETi-1 (BioCentury Extra For Wednesday, Oct. 22, 2003). In thisphase II-study, just as in the preceding phase I-study, a very goodsafety profile without any questionable side effects was proven,allowing the conclusion to be drawn that basically no side effects areto be expected from an anti-CETP vaccination approach. With regard toefficacy, however, the Avant vaccine was disappointing since it did notlead to increased HDL levels significantly better than those attained bya placebo treatment.

The problem with the CETi-1 vaccine is that it uses endogenous antigen.The human immune system is tolerant relative to endogenous structures,since with most of the endogenous molecules—other than with CETP—it isvital that no autoantibodies be formed. Thus, it was the object of theCETi-1 vaccine to break the endogenous tolerance which, apparently, ithas not achieved to a sufficient extent.

Thus, it is the object of the present invention to provide antigens foran anti-CETP vaccine which are selected such that they are considered asforeign by the immune system and therefore need not break aself-tolerance. These antigens may be used for preventing and/ortreating atherosclerosis, atherosclerosis risk diseases andatherosclerosis sequelae.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 shows the result of a representative competition ELISA afterscreening phage display library Ph.D. 7 with monoclonal antibody“Paula”.

FIGS. 2 a and 2 b show the results of 2 typical competition ELISAs afterscreening phage display library Ph.D. 12 with monoclonal antibody“Paula”.

FIGS. 3 a and 3 b show the results of 2 representative competitionELISAs after screening phage display library Ph.D. 7 with mAb Frida.

FIG. 4 a shows the result of a representative competition ELISA afterscreening phage display library Ph.D. 12 with monoclonal antibody“Frida”.

FIG. 4 b shows binding of monoclonal antibody “Frida” to ELISA platescoated with mimotope-BSA.

FIGS. 5 a and 5 b show the results of a representative competition ELISAafter screening phage display library Ph.D. 12 with monoclonal antibody“Frida”.

FIG. 6 shows the results of a competition ELISA of two mimotopes afterscreening phage display library Ph.D. 12 with monoclonal antibody“Frida”.

FIGS. 7 a, 7 b, 7 c and 7 d show the antibody titer (anti mouse IgG) ofin vivo experiments, whereby the following mimotope-BSA conjugates wereinjected into mice.

FIGS. 8 a and 8 b show the results of two representative competitionELISA after screening phage display library Ph.D. 7C7 with monoclonalantibody “Frida”.

FIG. 9 shows an in vitro ELISA test for the detection of the bindingbetween “Frida” and cyclic mimotopes.

FIGS. 10 a and 10 b show the results of an inhibition ELISA assay withFGFPSHLIIDWLQSLS (SEQ ID NO. 179), FGFPAHVFIDWLQSLS (SEQ ID NO. 222) andFGFPAHVYIDWLQSLS (SEQ ID NO. 223).

FIG. 11 shows the in vivo induction of antibodies directed to CETP bymimotopes of the invention that are administered to mice.

FIGS. 12 a and 12 b show the in vivo induction of CETP specificantibodies by the administration of the mimotopes of the invention.

FIG. 13 shows the in vivo induction of antibodies directed to CETP bymimotopes of the invention that are administered to mice.

FIG. 14 shows a CETP activity assay, wherein 0.6 μl human serum (withendogenous CETP activity) is mixed with serum from wild-type mice (notcontaining CETP activity) vaccinated with KLH/Alum (negative controlgroup), p4703-KLH/Alum (original CETP epitope), or p4361 (or p4362 or p4325) mimotope, respectively.

FIG. 15 shows that the addition of p4325-KLH/Alum to human seruminhibits significantly CETP activity.

FIG. 16 shows that the addition of p4361-KLH/Alum to human seruminhibits significantly CETP activity.

FIG. 17 shows that the addition of p4362-KLH/Alum to human seruminhibits significantly CETP activity.

FIG. 18 a shows an inhibition ELISA with mimotopes (Coat. 1 μM 4073peptide, detection α IgG1).

FIG. 18 b shows an inhibition ELISA with mimotopes (Coat. 1 μM 4073peptide, detection α IgG1).

FIG. 18 c shows a inhibition ELISA with mimotopes screen PhD12 Frida andAla-exchange for mimotope characterisation/mAb Frida (Coat 1 μM 4073.Detection αIgG1.)

FIG. 19 a shows a peptide ELISA, immunisation with C-DFGFPAHVYIDWLQSLS(p4628-KLH/Alum) (SEQ ID NO. 236), titre to original epitope.

FIG. 19 b shows a peptide ELISA, immunisation with C-FGFPAHVFIDWLQSLN(p4474-KLH/Alum) (SEQ ID NO. 230), titre to original epitope.

FIG. 19 c shows a peptide ELISA, immunisation with C-FGFPAHVFIDWLQSLN(p4474-KLH/Alum) (SEQ ID NO. 230), titre to injected mimotope.

FIG. 19 d shows an anti-protein ELISA. Mice were injected 3 times with30 μg of the indicated mimotopes coupled to KLH with Alum as adjuvant.Sera from each group (comprising 5 mice) were pooled, diluted 1:100 andtested on ELISA plates coated with purified rabbit CETP.

FIG. 19 e shows an anti-protein ELISA, wherein mice were injected 3times with 30 μg of the indicated mimotopes coupled to KLH with Alum asadjuvant. Mouse sera (from single mice) were diluted 1:100 and tested onELISA plates coated with purified rabbit CETP.

Therefore the present invention relates to the use of a compoundcomprising the amino acid sequence

(Z₁)_(n)X₁X₂X₃X₄(Z₂)_(m), (SEQ ID NO. 1)

whereinZ₁ is an amino acid residue other than C,X₁ is an amino acid residue selected from the group consisting of D, A,R, E, S, N, T and G,X₂ is an amino acid residue selected from the group consisting of F, A,W, R, S, L, Q, V and M,X₃ is an amino acid residue selected from the group consisting of L, A,S, W, E, R, I and H,X₄ is an amino acid residue selected from the group consisting of Q, A,H, D, K, R, S and E,Z₂ is an amino acid residue other than C,n is an integer between 0 and 10, preferably between 0 and 9,m is an integer between 0 and 3,is not, or does not comprise, a 4- to 16-mer polypeptide fragment of thecholesterol ester transport protein (CETP) or a CETP-epitope, saidcompound having a binding capacity to an antibody which is specific forthe natural CETP glycoprotein,orcomprising an amino acid sequence selected from the group consisting ofSYHATFL (SEQ ID NO. 2), TMAFPLN (SEQ ID NO. 3), HYHGAFL (SEQ ID NO. 4),EHHDIFL (SEQ ID NO. 5), TGLSVFL (SEQ ID NO. 6), WMPSLFY (SEQ ID NO. 7),SMPWWFF (SEQ ID NO. 8), TMPLLFW (SEQ ID NO. 9), DTWPGLE (SEQ ID NO. 10),SMPPIFY (SEQ ID NO. 11), MPLWWWD (SEQ ID NO. 12), SMPNLFY (SEQ ID NO.13), RMPPIFY (SEQ ID NO. 14), NPFEVFL (SEQ ID NO. 15), TLPNWFW (SEQ IDNO. 16), SMPLTFY (SEQ ID NO. 17), SPHPHFL (SEQ ID NO. 18), NFMSIGL (SEQID NO. 19), SQFLASL (SEQ ID NO. 20), WSWPGLN (SEQ ID NO. 21), IAWPGLD(SEQ ID NO. 22), SKFMDTL (SEQ ID NO. 23), SMPMVFY (SEQ ID NO. 24),YEWVGLM (SEQ ID NO. 25), KGFLDHL (SEQ ID NO. 26), HQSDDKMPWWFF (SEQ IDNO. 27), YVWQDPSFTTFF (SEQ ID NO. 28), YVWQDPSFTTFF (SEQ ID NO. 29),LPQTHPLHLLED (SEQ ID NO. 30), GPVSIYADTDFL (SEQ ID NO. 31), DSNDTLTLAAFL(SEQ ID NO. 32), NGSPALSHMLFL (SEQ ID NO. 33), TDYDPMWVFFGY (SEQ ID NO.34), IFPLDSQWQTFW (SEQ ID NO. 35), NESMPDLFYQPS (SEQ ID NO. 36),DWGDKYFSSFWN (SEQ ID NO. 37), VSAYNNV (SEQ ID NO. 38) and WPLHLWQ (SEQID NO. 39)for producing a medicament for preventing and/or treatingatherosclerosis, atherosclerosis risk diseases and atherosclerosissequelae.

The present invention provides CETP mimotopes for these purposes. Thesemimotopes are able to induce antibodies which are able to inhibit CETPenzyme activity. The CETP mimotopes according to the present inventionpreferably are antigenic polypeptides which in their amino acid sequencevary from the amino acid sequence of CETP or of fragments of CETP. Inthis respect, the inventive mimotopes may comprise one or morenon-natural amino acids (i.e. not from the 20 “classical” amino acids)or they may be completely assembled of such non-natural amino acids.Moreover, the inventive antigens which induce anti-CETP antibodies maybe assembled of D- or L-amino acids or of combinations of DL-amino acidsand, optionally, they may have been changed by further modifications,ring closures or derivatizations. Suitable anti-CETP-antibody-inducingantigens may be provided from commercially available peptide libraries.Preferably, these peptides are at least 4 amino acid residues in length,in particular at least 7 amino acids, and preferred lengths may be up to16, preferably up to 14 or 20 amino acids (e.g. 5 to 16 amino acidresidues). According to the invention, however, also longer peptides mayvery well be employed as anti-CETP-antibody-inducing antigens.Furthermore the mimotopes of the present invention may also be part of apolypeptide and consequently comprising at their N- and/or C-terminus atleast one further amino acid residue.

The mimotopes of the present invention are capable to bind to antibodieswhich may be obtained by administration of C-FGFPEHLLVDFLQSLS (SEQ IDNO. 146) (16 C-terminal amino acids of CETP protein) coupled to KLH orother carriers to mammals. Once administered to a mammal the mimotopesare able to induce a corresponding immune response, so that antibodiesdirected against CETP are produced in said mammal.

The CETP-mimotopes (i.e. anti-CETP-antibody-inducing antigens) of thepresent invention can be identified and prepared by various methods,including phage libraries or peptide libraries. They can be produced andidentified for instance by means of combinatorial chemistry or by meansof high throughput screening techniques for the most varying structures(Display: A Laboratory Manual by Carlos F. Barbas (Editor), et al.;Willats W G Phage display: practicalities and prospects. Plant Mol.Biol. 2002; 50(6):837-54).

Furthermore, according to the invention also anti-CETP-antibody-inducingantigens based on nucleic acids (“aptamers”) may be employed, and these,too, may be found with the most varying (oligonucleotide) libraries(e.g. with 2-180 nucleic acid residues) (e.g. Burgstaller et al., Curr.Opin. Drug Discov. Dev. 5(5) (2002), 690-700; Famulok et al., Acc. Chem.Res. 33 (2000), 591-599; Mayer et al., PNAS 98 (2001), 4961-4965, etc.).In anti-CETP-antibody-inducing antigens based on nucleic acids, thenucleic acid backbone can be provided e.g. by the naturalphosphor-diester compounds, or also by phosphorotioates or combinationsor chemical variations (e.g. as PNA), wherein as bases, according to theinvention primarily U, T, A, C, G, H and mC can be employed. The2′-residues of the nucleotides which can be used according to thepresent invention preferably are H, OH, F, Cl, NH₂, O-methyl, O-ethyl,O-propyl or O-butyl, wherein the nucleic acids may also be differentlymodified, i.e. for instance with protective groups, as they are commonlyemployed in oligonucleotide synthesis. Thus, aptamer-basedanti-CETP-antibody-inducing antigens are also preferredanti-CETP-antibody-inducing antigens within the scope of the presentinvention.

According to the present invention the term “mimotope” refers to amolecule which has a conformation that has a topology equivalent to theepitope of which it is a mimic. The mimotope binds to the sameantigen-binding region of an antibody which binds immunospecifically toa desired antigen. The mimotope will elicit an immunological response ina host that is reactive to the antigen to which it is a mimic. Themimotope may also act as a competitor for the epitope of which it is amimic in in vitro inhibition assays (e.g. ELISA inhibition assays) whichinvolve the epitope and an antibody binding to said epitope. However, amimotope of the present invention may not necessarily prevent or competewith the binding of the epitope of which it is a mimic in an in vitroinhibition assay although it is capable to induce a specific immuneresponse when administered to a mammal.

As used herein, the term “epitope” refers to an immunogenic region of anantigen which is recognized by a particular antibody molecule. Ingeneral, an antigen will possess one or more epitopes, each capable ofbinding an antibody that recognizes the particular epitope.

The abbreviations for the amino acid residues disclosed in the presentinvention follow the IUPAC recommendations:

Amino Acid 3-Letter Code 1-Letter Code Alanine Ala A Arginine Arg RAsparagine Asn N Aspartic Asp D Cysteine Cys C Glutamic Glu E GlutamineGln Q Glycine Gly G Histidine His H Isoleucine Ile I Leucine Leu LLysine Lys K Methionine Met M Phenylalanine Phe F Proline Pro P SerineSer S Threonine Thr T Tryptophan Trp W Tyrosine Tyr Y Valine Val V

The mimotopes of the present invention can be synthetically produced bychemical synthesis methods which are well known in the art, either as anisolated peptide or as a part of another peptide or polypeptide.Alternatively, the peptide mimotope can be produced in a microorganismwhich produces the peptide mimotope which is then isolated and ifdesired, further purified. The peptide mimotope can be produced inmicroorganisms such as bacteria, yeast or fungi, in eukaryote cells suchas a mammalian or an insect cells, or in a recombinant virus vector suchas adenovirus, poxvirus, herpesvirus, Simliki forest virus, baculovirus,bacteriophage, sindbis virus or sendai virus. Suitable bacteria forproducing the peptide mimotope include E. coli, B. subtilis or any otherbacterium that is capable of expressing peptides such as the peptidemimotope. Suitable yeast types for expressing the peptide mimotopeinclude Saccharomyces cerevisiae, Schizosaccharomyces pombe, Candida,Pichia pastoris or any other yeast capable of expressing peptides.Corresponding methods are well known in the art. Also methods forisolating and purifying recombinantly produced peptides are well knownin the art and include e.g. as gel filtration, affinity chromatography,ion exchange chromatography etc.

To facilitate isolation of the peptide mimotope, a fusion polypeptidemay be made wherein the peptide mimotope is translationally fused(covalently linked) to a heterologous polypeptide which enablesisolation by affinity chromatography. Typical heterologous polypeptidesare His-Tag (e.g. His₆; 6 histidine residues), GST-Tag(Glutathione-S-transferase) etc. The fusion polypeptide facilitates notonly the purification of the mimotopes but can also prevent the mimotopepolypeptide from being degraded during purification. If it is desired toremove the heterologous polypeptide after purification the fusionpolypeptide may comprise a cleavage site at the junction between thepeptide mimotope and the heterologous polypeptide. The cleavage siteconsists of an amino acid sequence that is cleaved with an enzymespecific for the amino acid sequence at the site (e.g. proteases).

The mimotopes of the present invention may also modified at or nearbytheir N- and/or C-termini so that at said positions a cysteine residueis bound thereto. In a preferred embodiment terminally positioned(located at the N- and C-termini of the peptide) cysteine residues areused to cyclize the peptides through a disulfide bond.

The mimotopes of the present invention may also be used in variousassays and kits, in particular in immunological assays and kits.Therefore, it is particularly preferred that the mimotope may be part ofanother peptide or polypeptide, particularly an enzyme which is used asa reporter in immunological assays. Such reporter enzymes include e.g.alkaline phosphatase or horseradish peroxidase.

The term “atherosclerosis sequelae” or “sequelae of atherosclerosis”refers to the diseases which are a consequence of atherosclerose. Thesediseases include among others peripheral arterial occlusive disease,coronary heart disease and apoplectic cerebral insultus (see e.g.Steinberg D. J. Lipid Res. (2005) 46: 179-190; Steinberg D et al. J.Lipid Res (2006) 47: 1339-1351).

According to another preferred embodiment of the present invention X₁ isD and X₄ is Q or H, preferably Q. Such a molecule preferably comprisesat its N-terminus further amino acid residues having the sequence X_(a)X_(b) X_(c) X_(d) X_(e) X_(f) (SEQ ID NO. 41), wherein X_(a) is P, Y, Tor K, X_(b) is an amino acid residue other than C, X_(c) is H, X_(d) isY, L, H, V, T, I or F, X_(e) is Y, I, P, L, Q, S, R, T, F or A and X_(f)is A, W, V, Q, L, S, I, R or T.

According to a preferred embodiment of the present invention n is 7, 8or 9, Z₁ is an amino acid residue other than C or selected from thegroup consisting of F, G, F, A, P, W, Y, S, G, D, L, E, K, T, P, I andM, preferably from the group consisting of F, G, F, A, P, Y, T, S, G, Kand D, and Z₂ is selected from the group consisting of S, L, A, W, L, N,T, I, Y and H.

According to a further preferred embodiment of the present invention X₁is selected from the group consisting of D, A, R, E and L, X₂ isselected from the group consisting of F, A, W, Q and R, X₃ is selectedfrom the group consisting of L, A and S, and X₄ is selected from thegroup consisting of Q, A and H.

According to a preferred embodiment of the present invention X₁ is D, X₂is selected from the group consisting of F, Q and W, X₃ is L or S and X₄is Q or H.

According to a preferred embodiment of the present invention thecompound comprises the amino acid sequence FX₈(F)_(o)PX₉HX₁₀X₁₁X₁₂DX₂X₃X₄X₅X₆X₇ (SEQ ID NO. 42), wherein

X₈ is selected from the group consisting of G, A, F, Y and K,X₉ is selected from the group consisting of E, Y, A, Q, K and S,X₁₀ is selected from the group consisting of H, V, L, F and I,X₁₁ is selected from the group consisting of L, W, S, I, F and Y,

X₁₂ is V, T, F or I, X₅ is S or Y, X₆ is L, A or I, X₇ is S, N or T, and

o is 0 or 1.

The compound of the present invention comprises preferably the aminoacid sequence X₁X₂X₃X₄X₅X₆X₇ (SEQ ID NO. 43), wherein X₁ is selectedfrom the group consisting of D, S, N, T and G, X₂ is F, X₃ is L, X₄ isselected from the group consisting of Q, D, K, R, S and E, X₅ is S or T,X₆ is L and X₇ is an amino acid residue other than C, preferablyselected from the group consisting of S, T, A, M, F and W.

According to a preferred embodiment of the present invention the aminoacid sequence is selected from the group consisting of SSLELFL (SEQ IDNO. 44), SFLDTLT (SEQ ID NO. 45), NFLKTLS (SEQ ID NO. 46), DFLRTLT (SEQID NO. 47), AFLDTLV (SEQ ID NO. 48), TFLSSLA (SEQ ID NO. 49), GFLDSLM(SEQ ID NO. 50), SPHPHFL (SEQ ID NO. 51), SNFLKTL (SEQ ID NO. 52),TGFLATL (SEQ ID NO. 53), SDFLRAL (SEQ ID NO. 54), SANPRDFLETLF (SEQ IDNO. 55), RMFPESFLDTLW (SEQ ID NO. 56), TIYDSFLDSLAS (SEQ ID NO. 57),KPYLLKDFLEAL (SEQ ID NO. 58), AMGPYDALDLFL (SEQ ID NO. 59), TWNPIESFLESL(SEQ ID NO. 60), QYQTPLTFLEAL (SEQ ID NO. 61), RHISPATFLEAL (SEQ ID NO.62), HTDSFLSTFYGD (SEQ ID NO. 63), ADSTFTSFLQTL (SEQ ID NO. 64),GPVSIYADTDFL (SEQ ID NO. 65), DSNDTLTLAAFL (SEQ ID NO. 66), TPTHYYADFSQL(SEQ ID NO. 67), LPGHLIWDSLHY (SEQ ID NO. 68), LPQTHPLHLLED (SEQ ID NO.69), IPYHHLVDQLHH (SEQ ID NO. 70), YPYHVQVDVLQN (SEQ ID NO. 71),IPSHHLQDSLQL (SEQ ID NO. 72), EYAHHTSLDLRQ (SEQ ID NO. 73), EPLHFRSDRIQA(SEQ ID NO. 74), ATPSHLIIDRAQ (SEQ ID NO. 75), APKHLYADMSQA (SEQ ID NO.76), FKPAHVSIDWLQ (SEQ ID NO. 77), MPAHLSRDLRQS (SEQ ID NO. 78),NPKHYSIDRHQA (SEQ ID NO. 79), SPQHLTTDRAQA (SEQ ID NO. 80), TPFHFAQDSWQW(SEQ ID NO. 81), TPTHYYADFSQLLS (SEQ ID NO. 82), TPTHYYADFSQSLS (SEQ IDNO. 83), GTPTHYYADFSQLL (SEQ ID NO. 84), GTPTHYYADFSQSL (SEQ ID NO. 85),FGTPTHYYADFSQSLS (SEQ ID NO. 86), FGFPTHYYADFSQSLS (SEQ ID NO. 87),LPGHLIWDSLHY (SEQ ID NO. 88), LPGHLIWDSLHYL (SEQ ID NO. 89),LPGHLIWDSLHYLS (SEQ ID NO. 90), LPGHLIWDSLHSL (SEQ ID NO. 91),LPGHLIWDSLHSLS (SEQ ID NO. 92), GLPGHLIWDSLHYL (SEQ ID NO. 93),GLPGHLIWDSLHSL (SEQ ID NO. 94), FGLPGHLIWDSLHSLS (SEQ ID NO. 95),FGFPGHLIWDSLHSLS (SEQ ID NO. 96), LPQTHPLHLLED (SEQ ID NO. 97),IPYHHLVDQLHH (SEQ ID NO. 98), IPYHHLVDQLHLS (SEQ ID NO. 99),IPYHHLVDQLHSLS (SEQ ID NO. 100), FGIPYHHLVDQLHHLS (SEQ ID NO. 101),FGFPYHHLVDQLHSLS (SEQ ID NO. 102), YPYHVQVDVLQN (SEQ ID NO. 103),YPYHVQVDVLQNLS (SEQ ID NO. 104), YPYHVQVDVLQSLS (SEQ ID NO. 105),FGYPYHVQVDVLQNLS (SEQ ID NO. 106), FGFPYHVQVDVLQSLS (SEQ ID NO. 107),IPSHHLQDSLQL (SEQ ID NO. 108), IPSHHLQDSLQLLS (SEQ ID NO. 109),IPSHHLQDSLQSLS (SEQ ID NO. 110), GIPSHHLQDSLQLL (SEQ ID NO. 111),FGIPSHHLQDSLQLLS (SEQ ID NO. 112), FGFPSHHLQDSLQSLS (SEQ ID NO. 113),EYAHHTSLDLRQ (SEQ ID NO. 114), EPLHFRSDRIQA (SEQ ID NO. 115),EPLHFRSDRIQALS (SEQ ID NO. 116), EPLHFRSDRIQSLS (SEQ ID NO. 117),GEPLHFRSDRIQAL (SEQ ID NO. 118), FGEPLHFRSDRIQALS (SEQ ID NO. 119),FGFPLHFRSDRIQSLS (SEQ ID NO. 120), APKHLYADMSQA (SEQ ID NO. 121),APKHLYADMSQALS (SEQ ID NO. 122), APKHLYADMSQSLS (SEQ ID NO. 123),GAPKHLYADMSQAL (SEQ ID NO. 124), FGFPKHLYADMSQSLS (SEQ ID NO. 125),MPAHLSRDLRQS (SEQ ID NO. 126), MPAHLSRDLRQSL (SEQ ID NO. 127),MPAHLSRDLRQSLS (SEQ ID NO. 128), GMPAHLSRDLRQSL (SEQ ID NO. 129),FGFPAHLSRDLRQSLS (SEQ ID NO. 130), NPKHYSIDRHQA (SEQ ID NO. 131),TPFHFAQDSWQW (SEQ ID NO. 132), TPFHFAQDSWQWLS (SEQ ID NO. 133),TPFHFAQDSWQSLS (SEQ ID NO. 134), GTPFHFAQDSWQWL (SEQ ID NO. 135),FGFPFHFAQDSWQSLS (SEQ ID NO. 136), ACSFAYLYRC (SEQ ID NO. 137),ACFMGDKWVC (SEQ ID NO. 138), ACVLYPKAIC (SEQ ID NO. 139), ACYMGQQFVC(SEQ ID NO. 140), ACLTAYLHWC (SEQ ID NO. 141), ACTLFPVAYC (SEQ ID NO.142), ACWLFPYAHC (SEQ ID NO. 143), ACKSINMWLC (SEQ ID NO. 144),ACQTINRWLC (SEQ ID NO. 145), FGFPEHLLVDFLQSLS (SEQ ID NO. 146),FGFPEHLLVDFLQSLS (SEQ ID NO. 147), FPEHLLVDFLQSL (SEQ ID NO. 148),AGFPEHLLVDFLQSLS (SEQ ID NO. 149), FAFPEHLLVDFLQSLS (SEQ ID NO. 150),FGAPEHLLVDFLQSLS (SEQ ID NO. 151), FGFAEHLLVDFLQSLS (SEQ ID NO. 152),FGFPAHLLVDFLQSLS (SEQ ID NO. 153), FGFPEALLVDFLQSLS (SEQ ID NO. 154),FGFPEHALVDFLQSLS (SEQ ID NO. 155), FGFPEHLAVDFLQSLS (SEQ ID NO. 156),FGFPEHLLADFLQSLS (SEQ ID NO. 157), FGFPEHLLVAFLQSLS (SEQ ID NO. 158),FGFPEHLLVDALQSLS (SEQ ID NO. 159), FGFPEHLLVDFAQSLS (SEQ ID NO. 160),FGFPEHLLVDFLASLS (SEQ ID NO. 161), FGFPEHLLVDFLQALS (SEQ ID NO. 162),FGFPEHLLVDFLQSAS (SEQ ID NO. 163), FGFPEHLLVDFLQSLA (SEQ ID NO. 164),FAFPAHLLVDFLQALA (SEQ ID NO. 165), AAFPAHLLADFLQALA (SEQ ID NO. 166),SPQHLTTDRAQA (SEQ ID NO. 167), SPQHLTTDRAQALS (SEQ ID NO. 168),SPQHLTTDRAQSLS (SEQ ID NO. 169), GSPQHLTTDRAQAL (SEQ ID NO. 170),FGFPQHLTTDRAQSLS (SEQ ID NO. 171), FGFPQHLTTDWAQSLS (SEQ ID NO. 172),FGFPQHLTTDRLQSLS (SEQ ID NO. 173), FGFPQHLTTDWLQSLS (SEQ ID NO. 174),ATPSHLIIDRAQ (SEQ ID NO. 175), ATPSHLIIDRAQSLS (SEQ ID NO. 176),FGFPSHLIIDRAQSLS (SEQ ID NO. 177), FGFPSHLIIDWAQSLS (SEQ ID NO. 178),FGFPSHLIIDWLQSLS (SEQ ID NO. 179), FGFPSHLIIDWSQSLS (SEQ ID NO. 180),FATPSHLIIDWLQSLS (SEQ ID NO. 181), FKPAHVSIDWLQ (SEQ ID NO. 182),FKPAHVSIDWLQSLS (SEQ ID NO. 183), FGFPAHVSIDWLQSLS (SEQ ID NO. 184),AGFPAHVSIDWLQSLS (SEQ ID NO. 185), FAFPAHVSIDWLQSLS (SEQ ID NO. 186),FGAPAHVSIDWLQSLS (SEQ ID NO. 187), FGFAAHVSIDWLQSLS (SEQ ID NO. 188),FGFPAHVSADWLQSLS (SEQ ID NO. 189), FGFPAHVSIDWLQALS (SEQ ID NO. 190),FGFPAHVSIDWLQSLA (SEQ ID NO. 191), FAFPAHVSIDWLQALA (SEQ ID NO. 192),FGFAAHVSIDWLQSLS (SEQ ID NO. 193), FGFFAHVSIDWLQSLS (SEQ ID NO. 194),FGFPAHVSIRWLQSLS (SEQ ID NO. 195), FGFPAHVSIEWLQSLS (SEQ ID NO. 196),FGFPAHVSIDWLNSLS (SEQ ID NO. 197), FGFPAHVSIDWLHSLS (SEQ ID NO. 198),AGFPAHVSIDWLQSLS (SEQ ID NO. 199), PGFPAHVSIDWLQSLS (SEQ ID NO. 200),WGFPAHVSIDWLQSLS (SEQ ID NO. 201), FAFPAHVSIDWLQSLS (SEQ ID NO. 202),FSFPAHVSIDWLQSLS (SEQ ID NO. 203), FYFPAHVSIDWLQSLS (SEQ ID NO. 204),FDFPAHVSIDWLQSLS (SEQ ID NO. 205), FGAPAHVSIDWLQSLS (SEQ ID NO. 206),FGFPAHVSIDWLQLLS (SEQ ID NO. 207), FGFPAHVSIDWLQWLS (SEQ ID NO. 208),FGFPAHVSIDWLQNLS (SEQ ID NO. 209), FGFPAHVSIDWLQTLS (SEQ ID NO. 210),FGFPAHVSIDWLQYLS (SEQ ID NO. 211), FGFPAHVSIDWLQSIS (SEQ ID NO. 212),FGFPAHVSIDWLQSLT (SEQ ID NO. 213), FGFPAHVSIDWLQSLY (SEQ ID NO. 214),FAFPAHVFIDWLQALA (SEQ ID NO. 215), FGFPAHVSIDRAQSLS (SEQ ID NO. 216),FGFPTHVSIDWLQSLS (SEQ ID NO. 217), FGFPFHVSIDWLQSLS (SEQ ID NO. 218),FGFPAHISIDWLQSLS (SEQ ID NO. 219), FGFPAHIIIDWLQSLS (SEQ ID NO. 220),FGFPAHLTTDWLQSLS (SEQ ID NO. 221), FGFPAHVFIDWLQSLS (SEQ ID NO. 222),FGFPAHVYIDWLQSLS (SEQ ID NO. 223), FGFPAHVSLDWLQSLS (SEQ ID NO. 224),FGFPAHVSADWLQSLS (SEQ ID NO. 225), TPTHYYADFSQSLS (SEQ ID NO. 226),FGFPAHVSIDWSQSLS (SEQ ID NO. 227), FGFPAHVSIDFSQSLS (SEQ ID NO. 228),FGFPSHIIIDWLQSLS (SEQ ID NO. 239), FGFPSHLIIEWLQSLS (SEQ ID NO. 240),AAFPAHLLADAAQALA (SEQ ID NO. 241), AAFPAHAAADFLQALA (SEQ ID NO. 242),AAFAAHLLADFLQAAA (SEQ ID NO. 243), AAAPAHLLVDAAQAAA (SEQ ID NO. 244),FAFPAHVFIDWLQSLS (SEQ ID NO. 245); FGFPAHVFIDWLQALS (SEQ ID NO. 246),FGFPAHVFIDWLQSLA (SEQ ID NO. 247), GFPAHVFIDWLQSLS (SEQ ID NO. 248),FPAHVFIDWLQSLS (SEQ ID NO. 249), PAHVFIDWLQSLS (SEQ ID NO. 250),FAFPAHVFIDWLQALA (SEQ ID NO. 251), FGFPEHLFVDFLQSLS (SEQ ID NO. 252),FGFPAHVHIDWLQSLS (SEQ ID NO. 253), FGFPAHVPIDWLQSLS (SEQ ID NO. 254),FGFPSHLFIDWAQSLS (SEQ ID NO. 255), PGFPAHVFIDWLQLIT (SEQ ID NO. 256),PAHVYIDWLQSLS (SEQ ID NO. 257), FGFPAHVYIDWLQ (SEQ ID NO. 258),FGFPAHVFIDWLQ (SEQ ID NO. 259), DFGFPSHLIIDWLQSLS (SEQ ID NO. 235),DFGFPAHVFIDWLQSLN (SEQ ID NO. 260), PSHLIIDWLQ (SEQ ID NO. 261),PAHVFIDWLQ (SEQ ID NO. 262), DFGFPAHVTIDWLQSLN (SEQ ID NO. 263),DFGFPAHVLIDWLQSLN (SEQ ID NO. 264), FGFPAHVFIDWLQSLN (SEQ ID NO. 230)and FGFPAHVFIDWLQSLA (SEQ ID NO. 265).

Particularly preferred mimotopes to be used according to the presentinvention are SANPRDFLETLF (SEQ ID NO. 55), RMFPESFLDTLW (SEQ ID NO.56), SFLDTLT (SEQ ID NO. 45), NFLKTLS (SEQ ID NO. 46), DFLRTLT (SEQ IDNO. 47), TFLSSLA (SEQ ID NO. 49), GFLDSLM (SEQ ID NO. 50),FGFPYHVQVDVLQSLS (SEQ ID NO. 107), FGFPSHLIIDRAQSLS (SEQ ID NO. 177),FKPAHVSIDWLQSLS (SEQ ID NO. 183), FGFPAHVSIDWLQSLS (SEQ ID NO. 184),FGFPQHLTTDRAQSLS (SEQ ID NO. 171), FGFPTHYYADFSQSLS (SEQ IN NO. 87),FGFPGHLIWDSLHSLS (SEQ ID NO. 96), FGFPYHHLVDQLHSLS (SEQ ID NO. 102),FGFPSHHLQDSLQSLS (SEQ ID NO. 113), FGFPLHFRSDRIQSLS (SEQ ID NO. 120),FGFPKHLYADMSQSLS (SEQ ID NO. 125), FGFPAHLSRDLRQSLS (SEQ ID NO. 130) andFGFPFHFAQDSWQSLS (SEQ ID NO. 136).

Especially preferred mimotopes of the present invention areFGFPSHLIIDWLQSLS (SEQ ID NO. 179), FGFPAHVFIDWLQSLS (SEQ ID NO. 222) andFGFPAHVYIDWLQSLS(SEQ ID NO. 223).

Further preferred mimotopes are FGFPAHVWIDWLQSLS (SEQ ID NO. 229),FGFPAHVFIDWLQSLN (SEQ ID NO. 230), FGFPAHFSIDWLQSLS (SEQ ID NO. 231),FGFPAHVSFDWLQSLS (SEQ ID NO. 232), FGFPEHVFIDWLQSLS (SEQ ID NO. 233),DFGFPAHVFIDWLQSLS (SEQ ID NO. 234), DFGFPSHLIIDWLQSLS (SEQ ID NO. 235),DFGFPAHVYIDWLQSLS (SEQ ID NO. 236), FGFPQHLFTDWLQSLS (SEQ ID NO. 237)and FGFPKHLLVDFLQSLS (SEQ ID NO. 238).

According to a preferred embodiment of the present invention thecompound is coupled to a pharmaceutically acceptable carrier, preferablyKLH (Keyhole Limpet Hemocyanin), tetanus toxoid, albumin-bindingprotein, bovine serum albumin, a dendrimer (MAP; Biol. Chem. 358: 581),peptide linkers (or flanking regions) as well as the adjuvant substancesdescribed in Singh et al., Nat. Biotech. 17 (1999), 1075-1081 (inparticular those in Table 1 of that document), and O'Hagan et al.,Nature Reviews, Drug Discovery 2 (9) (2003), 727-735 (in particular theendogenous immuno-potentiating compounds and delivery systems describedtherein), or mixtures thereof. The conjugation chemistry (e.g. viaheterobifunctional compounds such as GMBS and of course also others asdescribed in “Bioconjugate Techniques”, Greg T. Hermanson) in thiscontext can be selected from reactions known to the skilled man in theart. Moreover, the vaccine composition may be formulated with anadjuvant, preferably a low soluble aluminium composition, in particularaluminium hydroxide. Of course, also adjuvants like MF59 aluminiumphosphate, calcium phosphate, cytokines (e.g., IL-2, IL-12, GM-CSF),saponins (e.g., QS21), MDP derivatives, CpG oligos, LPS, MPL,polyphosphazenes, emulsions (e.g., Freund's, SAF), liposomes, virosomes,iscoms, cochleates, PLG microparticles, poloxamer particles, virus-likeparticles, heat-labile enterotoxin (LT), cholera toxin (CT), mutanttoxins (e.g., LTK63 and LTR72), microparticles and/or polymerizedliposomes may be used.

The compound of the present invention is preferably bound to the carrieror adjuvant via a linker, which is selected from the group consisting ofNHS-poly (ethylene oxide) (PEO) (e.g. NHS-PEO₄-maleimide).

A vaccine which comprises the present compound (mimotope) and thepharmaceutically acceptable carrier may be administered by any suitablemode of application, e.g. i.d., i.v., i.p., i.m., intranasally, orally,subcutaneously, etc. and in any suitable delivery device (O'Hagan etal., Nature Reviews, Drug Discovery 2 (9), (2003), 727-735). Thecompound of the present invention is preferably formulated forintravenous, subcutaneous, intradermal or intramuscular administration(see e.g. “Handbook of Pharmaceutical Manufacturing Formulations”,Sarfaraz Niazi, CRC Press Inc, 2004).

Typically, the vaccine contains the compound according to the inventionin an amount of from 0.1 ng to 10 mg, preferably 10 ng to 1 mg, inparticular 100 ng to 100 μg, or, alternatively, e.g. 100 fmol to 10μmol, preferably 10 pmol to 1 μmol, in particular 100 pmol to 100 nmol.Typically, the vaccine may also contain auxiliary substances, e.g.buffers, stabilizers etc.

Another aspect of the present invention relates to a peptide consistingof at least one amino acid sequence selected from the group consistingof SYHATFL (SEQ ID NO. 2), TMAFPLN (SEQ ID NO. 3), HYHGAFL (SEQ ID NO.4), EHHDIFL (SEQ ID NO. 5), SSLELFL (SEQ ID NO. 44), TGLSVFL (SEQ ID NO.6), WMPSLFY (SEQ ID NO. 7), SMPWWFF (SEQ ID NO. 8), TMPLLFW (SEQ ID NO.9), DTWPGLE (SEQ ID NO. 10), SMPPIFY (SEQ ID NO. 11), MPLWWWD (SEQ IDNO. 12), SMPNLFY (SEQ ID NO. 13), RMPPIFY (SEQ ID NO. 14), NPFEVFL (SEQID NO. 15), TLPNWFW (SEQ ID NO. 16), SMPLTFY (SEQ ID NO. 17), SFLDTLT(SEQ ID NO. 45), NFLKTLS (SEQ ID NO. 46), DFLRTLT (SEQ ID NO. 47),AFLDTLV (SEQ ID NO. 48), TFLSSLA (SEQ ID NO. 49), GFLDSLM (SEQ ID NO.50), SPHPHFL (SEQ ID NO. 51), NFMSIGL (SEQ ID NO. 19), SQFLASL (SEQ IDNO. 20), SNFLKTL (SEQ ID NO. 52), TGFLATL (SEQ ID NO. 53), WSWPGLN (SEQID NO. 21), IAWPGLD (SEQ ID NO. 22), SKFMDTL (SEQ ID NO. 23), SDFLRAL(SEQ ID NO. 54), SMPMVFY (SEQ ID NO. 24), YEWVGLM (SEQ ID NO. 25),KGFLDHL (SEQ ID NO. 26), SANPRDFLETLF (SEQ ID NO. 55), RMFPESFLDTLW (SEQID NO. 56), TIYDSFLDSLAS (SEQ ID NO. 57), HQSDDKMPWWFF (SEQ ID NO. 27),KPYLLKDFLEAL (SEQ ID NO. 58), AMGPYDALDLFL (SEQ ID NO. 59), TWNPIESFLESL(SEQ ID NO. 60), YVWQDPSFTTFF (SEQ ID NO. 28), QYQTPLTFLEAL (SEQ ID NO.61), RHISPATFLEAL (SEQ ID NO. 62), HTDSFLSTFYGD (SEQ ID NO. 63),YVWQDPSFTTFF (SEQ ID NO. 29), ADSTFTSFLQTL (SEQ ID NO. 64), GPVSIYADTDFL(SEQ ID NO. 65), DSNDTLTLAAFL (SEQ ID NO. 66), NGSPALSHMLFL (SEQ ID NO.33), TDYDPMWVFFGY (SEQ ID NO. 34), IFPLDSQWQTFW (SEQ ID NO. 35),NESMPDLFYQPS (SEQ ID NO. 36), DWGDKYFSSFWN (SEQ ID NO. 37), VSAYNNV (SEQID NO. 38), WPLHLWQ (SEQ ID NO. 39), TPTHYYADFSQL (SEQ ID NO. 67),LPGHLIWDSLHY (SEQ ID NO. 68), LPQTHPLHLLED (SEQ ID NO. 69), IPYHHLVDQLHH(SEQ ID NO. 70), YPYHVQVDVLQN (SEQ ID NO. 71), IPSHHLQDSLQL (SEQ ID NO.72), EYAHHTSLDLRQ (SEQ ID NO. 73), EPLHFRSDRIQA (SEQ ID NO. 74),ATPSHLIIDRAQ (SEQ ID NO. 75), APKHLYADMSQA (SEQ ID NO. 76), FKPAHVSIDWLQ(SEQ ID NO. 77), MPAHLSRDLRQS (SEQ ID NO. 78), NPKHYSIDRHQA (SEQ ID NO.79), SPQHLTTDRAQA (SEQ ID NO. 80), TPFHFAQDSWQW (SEQ ID NO. 81),TPTHYYADFSQLLS (SEQ ID NO. 82), TPTHYYADFSQSLS (SEQ ID NO. 83),GTPTHYYADFSQLL (SEQ ID NO. 84), GTPTHYYADFSQSL (SEQ ID NO. 85),FGTPTHYYADFSQSLS (SEQ ID NO. 86), FGFPTHYYADFSQSLS (SEQ ID NO. 87),LPGHLIWDSLHY (SEQ ID NO. 88), LPGHLIWDSLHYL (SEQ ID NO. 89),LPGHLIWDSLHYLS (SEQ ID NO. 90), LPGHLIWDSLHSL (SEQ ID NO. 91),LPGHLIWDSLHSLS (SEQ ID NO. 92), GLPGHLIWDSLHYL (SEQ ID NO. 93),GLPGHLIWDSLHSL (SEQ ID NO. 94), FGLPGHLIWDSLHSLS (SEQ ID NO. 95),FGFPGHLIWDSLHSLS (SEQ ID NO. 96), LPQTHPLHLLED (SEQ ID NO. 97),IPYHHLVDQLHH (SEQ ID NO. 98), IPYHHLVDQLHLS (SEQ ID NO. 99),IPYHHLVDQLHSLS (SEQ ID NO. 100), FGIPYHHLVDQLHHLS (SEQ ID NO. 101),FGFPYHHLVDQLHSLS (SEQ ID NO. 102), YPYHVQVDVLQN (SEQ ID NO. 103),YPYHVQVDVLQNLS (SEQ ID NO. 104), YPYHVQVDVLQSLS (SEQ ID NO. 105),FGYPYHVQVDVLQNLS (SEQ ID NO. 106), FGFPYHVQVDVLQSLS (SEQ ID NO. 107),IPSHHLQDSLQL (SEQ ID NO. 108), IPSHHLQDSLQLLS (SEQ ID NO. 109),IPSHHLQDSLQSLS (SEQ ID NO. 110), GIPSHHLQDSLQLL (SEQ ID NO. 111),FGIPSHHLQDSLQLLS (SEQ ID NO. 112), FGFPSHHLQDSLQSLS (SEQ ID NO. 113),EYAHHTSLDLRQ (SEQ ID NO. 114), EPLHFRSDRIQA (SEQ ID NO. 115),EPLHFRSDRIQALS (SEQ ID NO. 116), EPLHFRSDRIQSLS (SEQ ID NO. 117),GEPLHFRSDRIQAL (SEQ ID NO. 118), FGEPLHFRSDRIQALS (SEQ ID NO. 119),FGFPLHFRSDRIQSLS (SEQ ID NO. 120), APKHLYADMSQA (SEQ ID NO. 121),APKHLYADMSQALS (SEQ ID NO. 122), APKHLYADMSQSLS (SEQ ID NO. 123),GAPKHLYADMSQAL (SEQ ID NO. 124), FGFPKHLYADMSQSLS (SEQ ID NO. 125),MPAHLSRDLRQS (SEQ ID NO. 126), MPAHLSRDLRQSL (SEQ ID NO. 127),MPAHLSRDLRQSLS (SEQ ID NO. 128), GMPAHLSRDLRQSL (SEQ ID NO. 129),FGFPAHLSRDLRQSLS (SEQ ID NO. 130), NPKHYSIDRHQA (SEQ ID NO. 131),TPFHFAQDSWQW (SEQ ID NO. 132), TPFHFAQDSWQWLS (SEQ ID NO. 133),TPFHFAQDSWQSLS (SEQ ID NO. 134), GTPFHFAQDSWQWL (SEQ ID NO. 135),FGFPFHFAQDSWQSLS (SEQ ID NO. 136), ACSFAYLYRC (SEQ ID NO. 137),ACFMGDKWVC (SEQ ID NO. 138), ACVLYPKAIC (SEQ ID NO. 139), ACYMGQQFVC(SEQ ID NO. 140), ACLTAYLHWC (SEQ ID NO. 141), ACTLFPVAYC (SEQ ID NO.142), ACWLFPYAHC (SEQ ID NO. 143), ACKSINMWLC (SEQ ID NO. 144),ACQTINRWLC (SEQ ID NO. 145), FGFPEHLLVDFLQSLS (SEQ ID NO. 146),FGFPEHLLVDFLQSLS (SEQ ID NO. 147), FPEHLLVDFLQSL (SEQ ID NO. 148),AGFPEHLLVDFLQSLS (SEQ ID NO. 149), FAFPEHLLVDFLQSLS (SEQ ID NO. 150),FGAPEHLLVDFLQSLS (SEQ ID NO. 151), FGFAEHLLVDFLQSLS (SEQ ID NO. 152),FGFPAHLLVDFLQSLS (SEQ ID NO. 153), FGFPEALLVDFLQSLS (SEQ ID NO. 154),FGFPEHALVDFLQSLS (SEQ ID NO. 155), FGFPEHLAVDFLQSLS (SEQ ID NO. 156),FGFPEHLLADFLQSLS (SEQ ID NO. 157), FGFPEHLLVAFLQSLS (SEQ ID NO. 158),FGFPEHLLVDALQSLS (SEQ ID NO. 159), FGFPEHLLVDFAQSLS (SEQ ID NO. 160),FGFPEHLLVDFLASLS (SEQ ID NO. 161), FGFPEHLLVDFLQALS (SEQ ID NO. 162),FGFPEHLLVDFLQSAS (SEQ ID NO. 163), FGFPEHLLVDFLQSLA (SEQ ID NO. 164),FAFPAHLLVDFLQALA (SEQ ID NO. 165), AAFPAHLLADFLQALA (SEQ ID NO. 166),SPQHLTTDRAQA (SEQ ID NO. 167), SPQHLTTDRAQALS (SEQ ID NO. 168),SPQHLTTDRAQSLS (SEQ ID NO. 169), GSPQHLTTDRAQAL (SEQ ID NO. 170),FGFPQHLTTDRAQSLS (SEQ ID NO. 171), FGFPQHLTTDWAQSLS (SEQ ID NO. 172),FGFPQHLTTDRLQSLS (SEQ ID NO. 173), FGFPQHLTTDWLQSLS (SEQ ID NO. 174),ATPSHLIIDRAQ (SEQ ID NO. 175), ATPSHLIIDRAQSLS (SEQ ID NO. 176),FGFPSHLIIDRAQSLS (SEQ ID NO. 177), FGFPSHLIIDWAQSLS (SEQ ID NO. 178),FGFPSHLIIDWLQSLS (SEQ ID NO. 179), FGFPSHLIIDWSQSLS (SEQ ID NO. 180),FATPSHLIIDWLQSLS (SEQ ID NO. 181), FKPAHVSIDWLQ (SEQ ID NO. 182),FKPAHVSIDWLQSLS (SEQ ID NO. 183), FGFPAHVSIDWLQSLS (SEQ ID NO. 184),AGFPAHVSIDWLQSLS (SEQ ID NO. 185), FAFPAHVSIDWLQSLS (SEQ ID NO. 186),FGAPAHVSIDWLQSLS (SEQ ID NO. 187), FGFAAHVSIDWLQSLS (SEQ ID NO. 188),FGFPAHVSADWLQSLS (SEQ ID NO. 189), FGFPAHVSIDWLQALS (SEQ ID NO. 190),FGFPAHVSIDWLQSLA (SEQ ID NO. 191), FAFPAHVSIDWLQALA (SEQ ID NO. 192),FGFAAHVSIDWLQSLS (SEQ ID NO. 193), FGFFAHVSIDWLQSLS (SEQ ID NO. 194),FGFPAHVSIRWLQSLS (SEQ ID NO. 195), FGFPAHVSIEWLQSLS (SEQ ID NO. 196),FGFPAHVSIDWLNSLS (SEQ ID NO. 197), FGFPAHVSIDWLHSLS (SEQ ID NO. 198),AGFPAHVSIDWLQSLS (SEQ ID NO. 199), PGFPAHVSIDWLQSLS (SEQ ID NO. 200),WGFPAHVSIDWLQSLS (SEQ ID NO. 201), FAFPAHVSIDWLQSLS (SEQ ID NO. 202),FSFPAHVSIDWLQSLS (SEQ ID NO. 203), FYFPAHVSIDWLQSLS (SEQ ID NO. 204),FDFPAHVSIDWLQSLS (SEQ ID NO. 205), FGAPAHVSIDWLQSLS (SEQ ID NO. 206),FGFPAHVSIDWLQLLS (SEQ ID NO. 207), FGFPAHVSIDWLQWLS (SEQ ID NO. 208),FGFPAHVSIDWLQNLS (SEQ ID NO. 209), FGFPAHVSIDWLQTLS (SEQ ID NO. 210),FGFPAHVSIDWLQYLS (SEQ ID NO. 211), FGFPAHVSIDWLQSIS (SEQ ID NO. 212),FGFPAHVSIDWLQSLT (SEQ ID NO. 213), FGFPAHVSIDWLQSLY (SEQ ID NO. 214),FAFPAHVSIDWLQALA (SEQ ID NO. 215), FGFPAHVSIDRAQSLS (SEQ ID NO. 216),FGFPTHVSIDWLQSLS (SEQ ID NO. 217), FGFPFHVSIDWLQSLS (SEQ ID NO. 218),FGFPAHISIDWLQSLS (SEQ ID NO. 219), FGFPAHIIIDWLQSLS (SEQ ID NO. 220),FGFPAHLTTDWLQSLS (SEQ ID NO. 221), FGFPAHVFIDWLQSLS (SEQ ID NO. 222),FGFPAHVYIDWLQSLS (SEQ ID NO. 223), FGFPAHVSLDWLQSLS (SEQ ID NO. 224),FGFPAHVSADWLQSLS (SEQ ID NO. 225), TPTHYYADFSQSLS (SEQ ID NO. 226),FGFPAHVWIDWLQSLS (SEQ ID NO. 229), FGFPAHVFIDWLQSLN (SEQ ID NO. 230),FGFPAHFSIDWLQSLS (SEQ ID NO. 231), FGFPAHVSFDWLQSLS (SEQ ID NO. 232),FGFPEHVFIDWLQSLS (SEQ ID NO. 233), DFGFPAHVFIDWLQSLS (SEQ ID NO. 234),DFGFPSHLIIDWLQSLS (SEQ ID NO. 235), DFGFPAHVYIDWLQSLS (SEQ ID NO. 236),FGFPQHLFTDWLQSLS (SEQ ID NO. 237), FGFPKHLLVDFLQSLS (SEQ ID NO. 238),FGFPAHVSIDFSQSLS (SEQ ID NO. 227), FGFPAHVSIDFSQSLS (SEQ ID NO. 228),FGFPSHIIIDWLQSLS (SEQ ID NO. 239), FGFPSHLIIEWLQSLS (SEQ ID NO. 240),AAFPAHLLADAAQALA (SEQ ID NO. 241), AAFPAHAAADFLQALA (SEQ ID NO. 242),AAFAAHLLADFLQAAA (SEQ ID NO. 243), AAAPAHLLVDAAQAAA (SEQ ID NO. 244),FAFPAHVFIDWLQSLS (SEQ ID NO. 245); FGFPAHVFIDWLQALS (SEQ ID NO. 246),FGFPAHVFIDWLQSLA (SEQ ID NO. 247), GFPAHVFIDWLQSLS (SEQ ID NO. 248),FPAHVFIDWLQSLS (SEQ ID NO. 249), PAHVFIDWLQSLS (SEQ ID NO. 250),FAFPAHVFIDWLQALA (SEQ ID NO. 251), FGFPEHLFVDFLQSLS (SEQ ID NO. 252),FGFPAHVHIDWLQSLS (SEQ ID NO. 253), FGFPAHVPIDWLQSLS (SEQ ID NO. 254),FGFPSHLFIDWAQSLS (SEQ ID NO. 255), PGFPAHVFIDWLQLIT (SEQ ID NO. 256),PAHVYIDWLQSLS (SEQ ID NO. 257), FGFPAHVYIDWLQ (SEQ ID NO. 258),FGFPAHVFIDWLQ (SEQ ID NO. 259), DFGFPAHVFIDWLQSLN (SEQ ID NO. 260),PSHLIIDWLQ (SEQ ID NO. 261), PAHVFIDWLQ (SEQ ID NO. 262),DFGFPAHVTIDWLQSLN (SEQ ID NO. 263), DFGFPAHVLIDWLQSLN (SEQ ID NO. 264),and FGFPAHVFIDWLQSLA (SEQ ID NO. 265).

The peptides of the present invention turned out to be mimotopes forCETP and, hence, the mimotopes were able to bind to antibodies bindingto the CETP fragment C-FGFPEHLLVDFLQSLS (SEQ ID NO. 146) (16 C-terminalamino acids of CETP protein).

Yet, another aspect of the present invention relates to a pharmaceuticalformulation comprising at least one peptide according to the presentinvention.

The peptides of the present invention may be formulated in apharmaceutical formulation which may be administered to an individual.These formulations may be used, e.g., for preventing and/or treatingatherosclerosis, atherosclerosis risk diseases and atherosclerosissequelae.

The peptides in the formulation can be combined from the pool ofpeptides disclosed herein. Furthermore is also possible to providepharmaceutical formulations, which comprise one or more of the peptidesof the present invention, and which can be administered separately ortogether to an individual in need thereof.

The peptides of the present invention can be mixed into one singlepharmaceutical formulation or in a combination of two or three. Theresulting formulation can be administered at the same or the differentmoments in time. According to a preferred embodiment of the presentinvention the peptide present in the formulation is coupled to apharmaceutically acceptable carrier, preferably KLH (Keyhole LimpetHemocyanin).

The present invention is further illustrated by the following figuresand examples, however, without being restricted thereto.

FIG. 1 shows the result of a representative competition ELISA afterscreening phage display library Ph.D. 7 with monoclonal antibody“Paula”.

FGFPEHLLVDFLQSLS SEQ ID NO. 146 FGFPEHLLVDFLQSLS SEQ ID NO. 147 SYHATFLSEQ ID NO. 2 TMAFPLN SEQ ID NO. 3 HYHGAFL SEQ ID NO. 4 EHHDIFLSEQ ID NO. 5 SSLELFL SEQ ID NO. 44 TGLSVFL SEQ ID NO. 6 WMPSLFYSEQ ID NO. 7 SMPWWFF SEQ ID NO. 8 TMPLLFW SEQ ID NO. 9 DTWPGLESEQ ID NO. 10 SMPPIFY SEQ ID NO. 11 MPLWWWD SEQ ID NO. 12 SMPNLFYSEQ ID NO. 13 RMPPIFY SEQ ID NO. 14 NPFEVFL SEQ ID NO. 15 TLPNWFWSEQ ID NO. 16

FIGS. 2 a and 2 b show the results of 2 typical competition ELISAs afterscreening phage display library Ph.D. 12 with monoclonal antibody“Paula”.

FGFPEHLLVDFLQSLS SEQ ID NO. 146 FGFPEHLLVDFLQSLS SEQ ID NO. 147RHISPATFLEAL SEQ ID NO. 62 TIYDSFLDSLAS SEQ ID NO. 57 HTDSFLSTFYGDSEQ ID NO. 63 ADSTFTSFLQTL SEQ ID NO. 64 GPVSIYADTDFL SEQ ID NO. 65DSNDTLTLAAFL SEQ ID NO. 66 NGSPALSHMLFL SEQ ID NO. 33 TDYDPMWVFFGYSEQ ID NO. 34 IFPLDSQWQTFW SEQ ID NO. 35 NESMPDLFYQPS SEQ ID NO. 36DWGDKYFSSFWN SEQ ID NO. 37 HQSDDKMPWWFF SEQ ID NO. 27 KPYLLKDFLEALSEQ ID NO. 58 SANPRDFLETLF SEQ ID NO. 55 RMFPESFLDTLW SEQ ID NO. 56AMGPYDALDLFL SEQ ID NO. 59 TWNPIESFLESL SEQ ID NO. 60 YVWQDPSFTTFFSEQ ID NO. 29 QYQTPLTFLEAL SEQ ID NO. 61

FIGS. 3 a and 3 b show the results of 2 representative competitionELISAs after screening phage display library Ph.D. 7 with mAb Frida.

FGFPEHLLVDFLQSLS SEQ ID NO. 146 FGFPEHLLVDFLQSLS SEQ ID NO. 147 WPLHLWQSEQ ID NO. 39 VSAYNNV SEQ ID NO. 38

FIG. 4 a shows the result of a representative competition ELISA afterscreening phage display library Ph.D. 12 with monoclonal antibody“Frida”.

FGFPEHLLVDFLQSLS SEQ ID NO. 146 FGFPEHLLVDFLQSLS SEQ ID NO. 147TPTHYYADFSQL SEQ ID NO. 67 LPGHLIWDSLHY SEQ ID NO. 68 LPQTHPLHLLEDSEQ ID NO. 69

FIG. 4 b shows binding of monoclonal antibody “Frida” to ELISA platescoated with mimotope-BSA

FIGS. 5 a and 5 b show the results of a representative competition ELISAafter screening phage display library Ph.D. 12 with monoclonal antibody“Frida”.

FGFPEHLLVDFLQSLS SEQ ID NO. 146 FGFPEHLLVDFLQSLS SEQ ID NO. 147GTPTHYYADFSQLL SEQ ID NO. 84 GTPTHYYADFSQSL SEQ ID NO. 85FGTPTHYYADFSQSLS SEQ ID NO. 86 FGFPTHYYADFSQSLS SEQ ID NO. 87GLPGHLIWDSLHYL SEQ ID NO. 93 GLPGHLIWDSLHSL SEQ ID NO. 94FGLPGHLIWDSLHSLS SEQ ID NO. 95 FGFPGHLIWDSLHSLS SEQ ID NO. 96FGIPYHHLVDQLHHLS SEQ ID NO. 101 FGFPYHHLVDQLHSLS SEQ ID NO. 102FGYPYHVQVDVLQNLS SEQ ID NO. 106 FGFPYHVQVDVLQSLS SEQ ID NO. 107GIPSHHLQDSLQLL SEQ ID NO. 111 FGIPSHHLQDSLQLLS SEQ ID NO. 112FGFPSHHLQDSLQSLS SEQ ID NO. 113 GEPLHFRSDRIQAL SEQ ID NO. 118FGEPLHFRSDRIQALS SEQ ID NO. 119 FGFPLHFRSDRIQSLS SEQ ID NO. 120ATPSHLIIDRAQSLS SEQ ID NO. 176 FGFPSHLIIDRAQSLS SEQ ID NO. 177GAPKHLYADMSQAL SEQ ID NO. 124 FGFPKHLYADMSQSLS SEQ ID NO. 125FKPAHVSIDWLQSLS SEQ ID NO. 183 FGFPAHVSIDWLQSLS SEQ ID NO. 184GMPAHLSRDLRQSL SEQ ID NO. 129 FGFPAHLSRDLRQSLS SEQ ID NO. 130GSPQHLTTDRAQAL SEQ ID NO. 170 FGFPQHLTTDRAQSLS SEQ ID NO. 171GTPFHFAQDSWQWL SEQ ID NO. 135 FGFPFHFAQDSWQSLS SEQ ID NO. 136

FIG. 6 shows the results of a competition ELISA of two mimotopes afterscreening phage display library Ph.D. 12 with monoclonal antibody“Frida”.

FGFPEHLLVDFLQSLS SEQ ID NO. 146 FGFPEHLLVDFLQSLS SEQ ID NO. 147FGFPSHLIIDWAQSLS SEQ ID NO. 178 FGFPSHLIIDWLQSLS SEQ ID NO. 179

FIGS. 7 a to 7 d show the antibody titer (anti mouse IgG) of in vivoexperiments, whereby the following mimotope-BSA conjugates were injectedinto mice:

Fr12/3/26/65 ext4 C-FGFPYHVQ  (SEQ ID NO. 107) p4286 VDVLQSLSFr12/3/55 ext2 C-FGFPSHLIIDRA (SEQ ID NO. 177) p4294 QSLSFr12/3/55 ext2 W instead of R  (SEQ ID NO. 178) p4324 C-FGFPSHLIIDWAQSLSFr12/3/55 ext2 WL instead of   (SEQ ID NO. 179) p4325RAC-FGFPSHLIIDWLQSLS Fr12/3/84 ext2 C-FGFPAHVSIDWL  (SEQ ID NO. 184)p4298 QSLS Fr12/3/40 ext4 C-FGFPQHLTTDRA (SEQ ID NO. 171) p4302 QSLSFr12/2/6 ext6 C-FGFPTHYYADFS  (SEQ ID NO. 87) p4278 QSLSFr12/2/11 ext7 C-FGFPGHLIWDSL  (SEQ ID NO. 96) p4282  HSLSFr12/3/1/19/88 ext4 C-FGFPYHHL (SEQ ID NO. 102) p4284 VDQLHSLSFr12/3/68 ext5 C-FGFPSHHLQDSL (SEQ ID NO. 113) p4289 QSLSFr12/3/83 ext5 C-FGFPLHFRSDRI (SEQ ID NO. 120) p4292 QSLSFr12/3/63 ext4 C-FGFPKHLYADMS (SEQ ID NO. 125) p4296 QSLSFr12/3/47 ext4 C-FGFPAHLSRDL (SEQ ID NO. 130) p4300 RQSLFr12/3/35 ext4 C-FGFPFHFAQDSW (SEQ ID NO. 136) p4304 QSLS

FIGS. 8 a and 8 b show the results of two representative competitionELISA after screening phage display library Ph.D. 7C7 with monoclonalantibody “Frida”.

FGFPEHLLVDFLQSLS SEQ ID NO. 146 FGFPEHLLVDFLQSLS SEQ ID NO. 147ACSFAYLYRC SEQ ID NO. 137 ACYMGQQFVC SEQ ID NO. 140 ACLTAYLHWCSEQ ID NO. 141 ACTLFPVAYC SEQ ID NO. 142 ACWLFPYAHC SEQ ID NO. 143ACQTINRWLC SEQ ID NO. 145

FIG. 9 shows an in vitro ELISA test for the detection of the bindingbetween “Frida” and cyclic mimotopes.

FGFPEHLLVDFLQSLS SEQ ID NO. 147 ACSFAYLYRC SEQ ID NO. 137 ACYMGQQFVCSEQ ID NO. 140 ACLTAYLHWC SEQ ID NO. 141 ACTLFPVAYC SEQ ID NO. 142ACWLFPYAHC SEQ ID NO. 143 ACQTINRWLC SEQ ID NO. 145

FIGS. 10 a and 10 b show the results of an inhibition ELISA assay withFGFPSHLIIDWLQSLS (SEQ ID NO. 179), FGFPAHVFIDWLQSLS (SEQ ID NO. 222) andFGFPAHVYIDWLQSLS (SEQ ID NO. 223).

FIG. 10 a (Coat 1 μM peptide. Detection αIgG1)

2.5 ng mAb Frida Frida 2 μg 20 μg pept N° peptide peptide buffer only —1.05 0.96 p4073 original epitope C-FGFPEHLLVDFLQSLS 0.44 0.1 p1358irrelevant peptide irrelevant peptide 1.08 0.91 p4361 FGFPAHVFIDWLQSLSFr12/3/84 ext2 VSI

VFI 0.82 0.16 p4362 FGFPAHVYIDWLQSLS Fr12/3/84 ext2 VSI

VYI 0.75 0.15

FIG. 10 b (Coat 1 μM peptide. Detection αIgG1)

2.5 ng mAb Firda Frida 2 μg 20 μg pept N^(o) peptide peptide buffer only— 0.84 0.75 p4073 original epitope C-FGFPEHLLVDFLQSLS 0.64 0.15 p1358irrelevant peptide irrelevant peptide 0.88 0.77 p4325 FGFPSHLIIDWLQSLSFr12/3/55 ext2 RA

WL 0.42 0.1

FIG. 11 shows the in vivo induction of antibodies directed to CETP bymimotopes of the invention that are administered to mice. Balb/c mice/30μg Peptide, 2 injections in 2 week intervals. S3=2 weeks after 3rdinjection. Alum as adjuvant. Titers against original epitope (p4073)induced by injection of mimotopes. Well coating: 50 μl of 1 μM p4073-BSAor 1 μg/ml activated KLH. Detection: αIgG:

injected original irrelevant peptide- epitope-peptide- BSA BSA BSAgroup 1 KLH KLH 2.040 400  group 2 original epitope p4073-KLH 8.600 10group 3 C-FGFPQHLTTDWLQSLS p4369-KLH 14.000 12.900 10 (SEQ ID NO. 174)group 4 C-FGFPSHLIIDWAQSLS p4324-KLH 12.570 7.600 10 (SEQ ID NO. 178)group 5 C-FGFPSHLIIDWLQSLS p4325-KLH 2.930 1.820 10 (SEQ ID NO. 179)group 6 C-FGFPSHLIIDWSQSLS p4366-KLH 4.700 3.600 10 (SEQ ID NO. 180)group 7 C-FATPSHLIIDWLQSLS p4345-KLH 8.380 1.270 10 (SEQ ID NO. 181)group 8 C-FAFPAHVSIDWLQALA p4328-KLH 10.100 2.740 400  (SEQ ID NO. 186)group 9 C-PGFPAHVSIDWLQSLS p4340-KLH 18.100 15.640 10 (SEQ ID NO. 200)group C-WGFPAHVSIDWLQSLS p4341-KLH 10.350 5.500 10 10 (SEQ ID NO. 201)group C-FSFPAHVSIDWLQSLS p4342-KLH 4.620 1.610 10 11 (SEQ ID NO. 203)group C-FYFPAHVSIDWLQSLS p4343-KLH 5.580 2.900 10 12 (SEQ ID NO. 204)group C-FDFPAHVSIDWLQSLS p4344-KLH 12.200 3.580 10 13 (SEQ ID NO. 205)group C-FGFPAHVSIDWLQLLS p4347-KLH 12.000 9.160 10 14 (SEQ ID NO. 207)group C-FGFPAHVSIDWLQYLS p4351-KLH 2.950 2.400 10 15 (SEQ ID NO. 211)group C-FGFPAHVSIDWLQSIS p4352-KLH 19.680 12.070 10 16 (SEQ ID NO. 212)group C-FGFPAHVSIDWLQSLT p4353-KLH 11.200 8.650 10 17 (SEQ ID NO. 213)group C-FGFPAHISIDWLQSLS p4358-KLH 16.500 12.940 10 18 (SEQ ID NO. 219)group C-FGFPAHIIIDWLQSLS p4359-KLH 8.540 5.340 10 19 (SEQ ID NO. 220)group C-FGFPAHVFIDWLQSLS p4361-KLH 17.940 9.530 10 20 (SEQ ID NO. 222)

FIGS. 12 a and 12 b show the in vivo induction of CETP specificantibodies by the administration of the mimotopes of the invention.Titers to p4073 and its correlation to titers to CETP of selected groups(which show high titers against p4073): gr.4, gr.9, gr.10, gr.14,gr.16-20/gr.1 (KLH), gr.2 (original epitope) as controls. Coating:recombinant GST-CETP or purified rabbit CETP, respectively:

FIG. 12 a

recombinant rabbit GST-CETP CETP group 1 KLH KLH/ 0.35 0.19 Alum group 2original epitope p4073- 1.49 1.25 KLH/ Alum group 3 C-FGFPQHLTTDWLQSLSp4369- 0.45 0.21 (SEQ ID NO. 174) KLH/ Alum group 4 C-FGFPSHLIIDWAQSLSp4324- 0.58 0.28 (SEQ ID NO. 178) KLH/ Alum group 9 C-PGFPAHVSIDWLQSLSp4340- 0.49 0.21 (SEQ ID NO. 200) KLH/ Alum group 10 C-WGFPAHVSIDWLQSLSp4341- 0.39 0.18 (SEQ ID NO. 201) KLH/ Alum group 14 C-FGFPAHVSIDWLQLLSp4347- 0.35 0.2 (SEQ ID NO. 207) KLH/ Alum group 16 C-FGFPAHVSIDWLQSISp4352- 0.48 0.28 (SEQ ID NO. 212) KLH/ Alum group 17 C-FGFPAHVSIDWLQSLTp4353- 0.57 0.39 (SEQ ID NO. 213) KLH/ Alum group 18 C-FGFPAHISIDWLQSLSp4358- 0.68 0.58 (SEQ ID NO. 219) KLH/ Alum group 19 C-FGFPAHIIIDWLQSLSp4359- 0.79 0.54 (SEQ ID NO. 220) KLH/ Alum group 20 C-FGFPAHVFIDWLQSLSp4361- 1.64 1.51 (SEQ ID NO. 222) KLH/ AlumFIG. 12 b

recombinant rabbit GST-CETP CETP group 1 KLH KLH/ 0.18 0.47 Alum group 2original epitope p4073- 1.26 1.42 KLH/ Alum group 5 C-FGFPSHLIIDWLQSLSp4325- 0.59 0.85 (SEQ ID NO. 179) KLH/ Alum group 6 C-FGFPSHLIIDWSQSLSp4366- 0.4 0.65 (SEQ ID NO. 180) KLH/ Alum group 7 C-FATPSHLIIDWLQSLSp4345- 0.39 0.46 (SEQ ID NO. 181) KLH/ Alum group 8 C-FAFPAHVSIDWLQALAp4328- 0.45 0.43 (SEQ ID NO. 186) KLH/ Alum group 11 C-FSFPAHVSIDWLQSLSp4342- 0.38 0.41 (SEQ ID NO. 203) KLH/ Alum group 12 C-FYFPAHVSIDWLQSLSp4343- 0.61 1.05 (SEQ ID NO. 204) KLH/ Alum group 13 C-FDFPAHVSIDWLQSLSp4344- 0.35 0.43 (SEQ ID NO. 205) KLH/ Alum group 15 C-FGFPAHVSIDWLQYLSp4351- 0.54 0.59 (SEQ ID NO. 211) KLH/ Alum

FIG. 13 shows the in vivo induction of antibodies directed to CETP bymimotopes of the invention that are administered to mice.

Sera of each group (5 Balb/c mice each) were combined, diluted 1:100 andtested on ELISA plates coated with recombinant GST-CETP or rabbit CETP,respectively. Detection of bound antibodies was with algG.

recombinant rabbit GST-CETP CETP group 1 KLH KLH/ 0.23 0.17 Alum group 2original epitope p4073- 1.08 0.46 KLH/ Alum group 3 C-FGFAAHVSIDWLQSLSp4335- 0.26 0.14 (SEQ ID NO. 188) KLH/ Alum group 4 C-FGFPAHVSIDWLQWLSp4348- 0.33 0.16 (SEQ ID NO. 208) KLH/ Alum group 5 C-FGFPAHLTTDWLQSLSp4360- 0.4 0.23 (SEQ ID NO. 221) KLH/ Alum group 6 C-FGFPAHVYIDWLQSLSp4362- 0.86 0.94 (SEQ ID NO. 223) KLH/ Alum group 7 C-FGFPAHVSIDWLQSLYP4354- 0.29 0.23 (SEQ ID NO. 214) KLH/ Alum group 8 C-FGFPAHVSIRWLQSLSp4337- 0.24 0.14 (SEQ ID NO. 195) KLH/ Alum

FIG. 14 shows a CETP activity assay, wherein 0.6 μl human serum (withendogenous CETP activity) is mixed with serum from wild-type mice (notcontaining CETP activity) vaccinated with KLH/Alum (negative controlgroup), p4703-KLH/Alum (original CETP epitope), or p4361 (or p4362 or p4325) mimotope, respectively. It could be demonstrated that the additionof 1.2 μl and 0.6 μl serum from p4361-KLH/Alum vaccinated micecompletely inhibits CETP activity and the addition of 0.2 μl serumreduces significantly said activity in contrast to the addition of serumfrom mice vaccinated with KLH/Alum-control only or with the originalepitope (p4073-KLH/Alum).

SEQ ID NO. 146 FGFPEHLLVDFLQSLS SEQ ID NO. 222 FGFPAHVFIDWLQSLS

FIG. 15 shows that the addition of p4325-KLH/Alum to human seruminhibits significantly CETP activity.

SEQ ID NO. 179 FGFPSHLIIDWLQSLS

FIG. 16 shows that the addition of p4361-KLH/Alum to human seruminhibits significantly CETP activity.

SEQ ID NO. 146 FGFPEHLLVDFLQSLS SEQ ID NO. 222 FGFPAHVFIDWLQSLS

FIG. 17 shows that the addition of p4362-KLH/Alum to human seruminhibits significantly CETP activity.

SEQ ID NO. 146 FGFPEHLLVDFLQSLS SEQ ID NO. 221 FGFPAHLTTDWLQSLSSEQ ID NO. 223 FGFPAHVYIDWLQSLS

FIG. 18 a shows an inhibition ELISA with mimotopes (Coat. 1 μM 4073peptide, detection α IgG1).

Frida 2.5 ng mAB Frida pept N^(o) low high buffer only buffer onlybuffer only 1.084 1.079 4% DMSO 4% DMSO 4% DMSO 1.180 1.201 p4073C-FGFPEHLLVDFLQSLS p4073 0.537 0.094 (SEQ ID NO. 147), positive controlpeptide p1208 positive control p1208 0.712 0.093 peptide FGFPEH-LLVDFLQSLS-C (SEQ ID NO. 146) p1358 negative control p1358 1.158 1.050peptide p4474 C-PAVVYIDWLQSLS Fr12/3/84 ext2 1.452 0.179(SEQ ID NO. 257) VSI

VFI SLS

SLN p4475 C-FGFPAHFSIDWLQSLS Fr12/3/84 ext2 2.211 1.429 (SEQ ID NO. 231)VSI

FSI p4476 C-FGFPAHVSFDWLQSLS Fr12/3/84 ext2 2.000 1.417 (SEQ ID NO. 232)VSI

VSF p4477 C-FGFPEHVFIDWLQSLS Fr12/3/84 ext2 0.808 0.116 (SEQ ID NO. 233)VSI

VFI PAH

PEH p4478 C-FKPAHVFIDWLQSLS Fr12/3/84 ext1 2.231 1.206 (SEQ ID NO. 266)VSI

VFI p4479 C-GFKPAHVFIDWLQSLS Fr12/3/84 ext1 2.165 1.591 (SEQ ID NO. 267)VSI

VFI plus G on N-terminus p4480 C-DFGPAHVFIDWLQSLS Fr12/3/84 ext2 0.5210.103 (SEQ ID NO. 234) VSI

VFI plus D on N-terminus; = 4361 plus D p4481 C-FGFPQHLFTDWLQSLSFr12/3/40 ext4 0.551 0.156 (SEQ ID NO. 237) RA

WL LTT

LFT = p4369 with exchange T

F

FIG. 18 b shows an inhibition ELISA with mimotopes (Coat. 1 μM 4073peptide, detection α IgG1).

p1208 positive control p1208 0.264 0.079 peptide p1358 negative controlp1358 1.902 1.661 peptide p4629 C-PAHVYIDWLQSLS C-terminus of p4362;0.313 0.118 (SEQ ID NO. 257) p4362 minus 3 aa on N-terminus p4630C-FGFPAHVYIDWLQ N-terminus of p4362 2.131 2.115 (SEQ ID NO. 258)(minus 3 aa on C-terminus) p4631 C-FGFPAHVFIDWLQ N-terminus of p43612.111 2.147 (SEQ ID NO. 259) (minus 3 aa on C-terminus) p4642C-DFGFPHSHLIIDWLQSLS Fr12/3/55 ext2 RA → 0.171 0.082 (SEQ ID NO. 235)WL plus D; p4325 plus D on N-terminus p4818 C-DFGFPAHVFIDWLQSLNFR12/3/84 ext 2 VSI → 0.332 0.091 (SEQ ID NO. 260) VFI SLS →SLN plus D; = 4361 N hinten plus D vorne p4819 C-PSHLIIDWLQ =4325 minus 3AA am N 2.226 2.158 (SEQ ID NO. 261) und am C-Terminus p4820C-PAHVFIDWLQ = 4361 minus 3AA am N 2.310 2.374 (SEQ ID NO. 262)und am C-Terminus p4989 C-DFGFPAHVTIDWLQSLN Fr12/3/84 ext2 VSI → 0.9320.274 (SEQ ID NO. 263) VTI; = p4361 F replaced by T,plus D on N-term and N instead of S on C-term p4990 C-DFGFPAHVLIDWLQSLNFr12/3/84 ext2 VSI → 0.263 0.073 (SEQ ID NO. 264) VLI; =p4361 F replaced by L, plus D on N-term and N instead of S on C-termp5067 FGFPAHVYIDWLQSLS-C p4362 C on C-terminus 0.563 0.217(SEQ ID NO. 223) p5068 FGFPAHVFIDWLQSLN-C p4474 C on C-terminus 0.7570.271 (SEQ ID NO. 230)

FIG. 18 c shows a inhibition ELISA with mimotopes screen PhD12 Frida andAla-exchange for mimotope characterisation/mAb Frida (Coat 1 μM 4073.Detection αIgG1.)

Frida 2.5 ng mAb Frida pept N^(o) low high buffer only buffer only 0.9640.964 4% DMSO 4% DMSO 0.973 0.923 positive control p4073 0.554 0.088peptide p1208 p1208 0.942 0.101 negative control p1358 0.986 0.93peptide p4432 C-FGFPSHIIIDWLQSLS Fr12/3/55 ext2exch2 0.635 0.096(SEQ ID NO. 239) L −> I p4433 C-FGFPSHLIIEWLQSLS Fr12/3/55 ext2exch21.114 0.672 (SEQ ID NO. 240) D −> E p4434 C-AAFPAHLLADAAQALAAla-exchange for 1.74 1.461 (SEQ ID NO. 241) mimotope characterisationp4435 C-AAFPAHAAADFLQALA Ala-exchange for 1.281 1.969 (SEQ ID NO. 242)mimotope characterisation p4436 C-AAFAAHLLADFLQAAA Ala-exchange for1.632 1.691 (SEQ ID NO. 243) mimotope characterisation p4437C-AAAPAHLLVDAAQAAA Ala-exchange for 1.84 1.674 (SEQ ID NO. 244)mimotope characterisation

FIG. 19 a shows a peptide ELISA, immunisation with C-DFGFPAHVYIDWLQSLS(p4628-KLH/Alum) (SEQ ID NO. 236), titre to original epitope.

FIG. 19 b shows a peptide ELISA, immunisation with C-FGFPAHVFIDWLQSLN(p4474-KLH/Alum) (SEQ ID NO. 230), titre to original epitope.

FIG. 19 c shows a peptide ELISA, immunisation with C-FGFPAHVFIDWLQSLN(p4474-KLH/Alum) (SEQ ID NO. 230), titre to injected mimotope.

FIG. 19 d shows an anti-protein ELISA. Mice were injected 3 times with30 μg of the indicated mimotopes coupled to KLH with Alum as adjuvant.Sera from each group (comprising 5 mice) were pooled, diluted 1:100 andtested on ELISA plates coated with purified rabbit CETP.

SEQ ID NO. 245 FAFPAHVFIDWLQSLS SEQ ID NO. 246 FGFPAHVFIDWLQALSSEQ ID NO. 247 FGFPAHVFIDWLQSLA SEQ ID NO. 248 GFPAHVFIDWLQSLSSEQ ID NO. 249 FPAHVFIDWLQSLS SEQ ID NO. 250 PAHVFIDWLQSLSSEQ ID NO. 251 FAFPAHVFIDWLQALA

FIG. 19 e shows an anti-protein ELISA, wherein mice were injected 3times with 30 μg of the indicated mimotopes coupled to KLH with Alum asadjuvant. Mouse sera (from single mice) were diluted 1:100 and tested onELISA plates coated with purified rabbit CETP.

SEQ ID NO. 252 FGFPEHLFVDFLQSLS SEQ ID NO. 253 FGFPAHVHIDWLQSLSSEQ ID NO. 254 FGFPAHVPIDWLQSLS SEQ ID NO. 269 FGFPAHVWIDWLQSLSEQ ID NO. 255 FGFPSHLFIDWAQSLS SEQ ID NO. 268 FPGFPAHVFIDWLQLIT

EXAMPLES

There exists a strong inverse relationship between the plasmaconcentration of cholesterol in high density lipoproteins (HDLs) and thedevelopment of coronary heart disease (CHD). Thus, the risk for CHD ishigher when HDLs decrease. Although 33% of patients with CHD have lowplasma levels of HDLs, there is currently no effective therapy forincreasing the plasma concentration of HDLs. Diet and moderate exerciseare ineffective, statins only achieve a low 5 to 7% increase in HDL, andniacin has side efects and compliance profiles limiting its use.

The inhibition of CETP activity has been suggested as therapeuticapproach to increase plasma HDL levels. CETP is a plasma glycoproteinthat facilitates transfer of neutral lipids and phospholipids betweenlipoproteins and regulates the concentration of plasma HDL. Theinhibition of CETP activity is expected to increase plasma HDLconcentrations for several reasons. CETP lowers HDL concentrations bymoving cholesteryl esters from HDLs to VLDLs and LDLs. Transientinhibition of CETP in rabbits and hamsters by monoclonal antibodies,small molecules (Sikorski, J. A., J. Med. Chem. 49 (1) (2006): 1-22), orantisense oligonucleotides causes HDL increase. Sustained CETPinhibition with antisense nucleotides increased plasma HDL and reducedatherosclerotic lesions in a rabbit model of atherosclerosis.CETP-transgenic mice and rats show decreased plasma HDL. Humans withreduced CETP activity have elevated plasma HDL.

Recently, a vaccine approach has been proposed. Rabbits were immunizedwith a human CETP-derived peptide containing a region of CETP criticalfor neutral lipid transfer function. Vaccinated rabbits had reduced CETPactivity and an altered lipoprotein profile with lower LDL and higherHDL concentration. Furthermore, CETP-vaccinated rabbits were shown tohave smaller atherosclerotic lesions than control animals.

The problem of the anti-CETP vaccine approach discussed above is thatthe vaccine formulation comprises a self peptide and therefore mustbreak natural tolerance against self antigens. The invention describes aCETP mimotope that can be used for vaccination: The mimotope shallinduce the production of antibodies against CETP. The CETP mimotope doesnot have a self sequence and therefore does not need to break tolerance.Thus, the induction of an anti-CETP antibody response is greatlyfacilitated. The mimotope is identified with a monoclonal antibody (mAb)and (commercially available) peptide libraries. An anti-CETP monoclonalantibody is used that neutralizes CETP activity. This mAb detects asequence within the C-terminal 26 amino acids of CETP necessary forneutral lipid transfer activity.

Example 1 Generation of Monoclonal Antibodies to be Used for Screeningof Phage Display Libraries

A.) 2 Antibodies Derived from “Fusion F”:

Balb/c mouse were immunized with original CETP epitopeC-FGFPEHLLVDFLQSLS (SEQ ID NO. 147) (16 C-terminal amino acids of CETPprotein) coupled to KLH and Alum as adjuvant.

2 hybridoma clones (both IgG1) were purified and used for screening:F5AF9G4 (“Paula”) and F6F11D1 (“Felix”).

These 2 monoclonal antibodies recognize the injected epitope as well asCETP protein in ELISA. They can also be used in Western Blot to detectCETP protein (recombinant protein expressed in bacteria as well asprotein isolated from rabbit serum). Both antibodies do not inhibit CETPenzyme activity (tested with Roar CETP Activity Assay Kit, see e.g. U.S.Pat. No. 5,585,235; U.S. Pat. No. 5,618,683; U.S. Pat. No. 5,770,355).

B.) 2 Antibodies Derived from “Fusion I”:

Balb/c mouse were immunized with original CETP epitopeC-FGFPEHLLVDFLQSLS (SEQ ID NO. 147) (16 C-terminal amino acids of CETPprotein) coupled to KLH and Alum as adjuvant.

2 hybridoma clones (both IgG1) were purified and used for screening:12G6H5 (“Frida”) and 12G6H7 (“James”).

These 2 monoclonal antibodies recognize the injected epitope as well asCETP protein in ELISA. They can also be used in Western Blot to detectCETP protein (recombinant protein expressed in bacteria as well asprotein isolated from rabbit serum). In contrast to the antibodiesderived from “Fusion F” (see A.)) both antibodies “Frida” and “James”inhibit CETP enzyme activity (tested with Roar CETP Activity Assay Kit).

Example 2 Phage Display, In Vitro Inhibition ELISA and In Vivo Testingof Mimotopes

Phage Display libraries used in this example were:

Ph.D. 7: New England BioLabs E8102L (linear 7mer library)

Ph.D. C7C: New England BioLabs E8121L (7mer library, cyclized peptides)

Ph.D. 12: New England BioLabs E8111L (linear 12mer library)

Phage Display was done according to manufacturer's protocol(www.neb.com).

After 2 or 3 subsequent rounds of panning, single phage clones werepicked and phage supernatants were subjected to ELISA on plates coatedwith the antibody that was used for the panning procedure. Phage clonesthat were positive in this ELISA (strong signal for the target, but nosignal for unspecific control) were sequenced. From DNA sequences,peptide sequences were deduced. These peptides were synthesized andcharacterised in inhibition ELISA.

1. In Vitro Inhibition Assay (ELISA)

Different amounts of peptides (2 and 20 μg, as indicated in therespective figures) derived from Phage Display were incubated with themonoclonal antibody that was used for the screening procedure. Peptidesdiminishing subsequent binding of the antibody to the original CETPepitope (C-terminal 16 amino acids of CETP protein) coated on ELISAplates were considered as inhibiting. (Results see i.a. FIGS. 19 a to 19c)

2. In Vivo Testing of Mimotopes

Inhibiting as well as some non-inhibiting peptides were coupled to KLHand injected into mice (wildtype or CETP-transgenic mice; subcutaneouslyinto the flank or intra-dermaly into the ears) or rabbits(subcutaneously into the flank) together with an appropriate adjuvant(aluminium hydroxide and Gerbu 100 for mice and aluminium hydroxide orCFA/IFA for rabbits).

Titers to injected peptides as well as to the original CETP epitope weredetermined. In addition, for selected sera also immune response to CETPprotein was measured (Results see FIGS. 7 a to 7 d and FIGS. 19 a to 19e).

3. Results

3.1. Screening with 2 Antibodies Derived from “Fusion F”: “Paula” and“Felix”

3.1.1. Phage Display Library Ph.D. 7

3.1.1.1. Screening with Monoclonal Antibody “Paula”

17 Sequences were identified in this screen:

(SEQ ID NO. 2) P2_8 SYHATFL (SEQ ID NO. 3) P2_9 TMAFPLN  (SEQ ID NO. 4)P2_11 HYHGAFL (SEQ ID NO. 5) P2_12 EHHDIFL (SEQ ID NO. 44) P2_15 SSLELFL(SEQ ID NO. 6) P2_16 TGLSVFL (SEQ ID NO. 7) P3_2 WMPSLFY (SEQ ID NO. 8)P3_6, 14, 28 SMPWWFF (SEQ ID NO. 9) P3_9 TMPLLFW (SEQ ID NO. 10) P3_13DTWPGLE (SEQ ID NO. 11) P3_16 SMPPIFY (SEQ ID NO. 12) P3_17 MPLWWWD(SEQ ID NO. 13) P3_18 SMPNLFY (SEQ ID NO. 14) P3_19 RMPPIFY(SEQ ID NO. 15) P3_21 NPFEVFL (SEQ ID NO. 16) P3_25 TLPNWFW(SEQ ID NO. 17) P3_26 SMPLTFY

The result of a representative competition ELISA is shown in FIG. 1.

3.1.1.2. Screening with Monoclonal Antibody “Felix”

6 sequences were identified that inhibit binding of monoclonal antibody“Felix” in in vitro competition experiments:

F2-9 C SFLDTLT (SEQ ID NO. 45) F3-6 C NFLKTLS (SEQ ID NO. 46) F3-18C DFLRTLT (SEQ ID NO. 47) F3-23 C AFLDTLV (SEQ ID NO. 48) F3-34C TFLSSLA (SEQ ID NO. 49) F3-38 C GFLDSLM (SEQ ID NO. 50)

Additional 12 sequences were identified that do not inhibit binding ofmonoclonal antibody “Felix” in in vitro competition experiments:

F2-2 + 5 SPHPHFL (SEQ ID NO. 51) F2-6 NFMSIGL (SEQ ID NO. 19)F2-16/F3-30 SQFLASL (SEQ ID NO. 20) F2-29 SNFLKTL (SEQ ID NO. 52) F3-1-_TGFLATL (SEQ ID NO. 53) F3-11-_ WSWPGLN (SEQ ID NO. 21) F3-17- IAWPGLD(SEQ ID NO. 22) F3-32- SKFMDTL (SEQ ID NO. 23) F3-41- SDFLRAL(SEQ ID NO. 54) F3-44-_ SMPMVFY (SEQ ID NO. 24) F3-49- YEWVGLM(SEQ ID NO. 25) F3-64- KGFLDHL (SEQ ID NO. 26

All mimotopes inhibiting the binding of monoclonal antibody “Felix” invitro were coupled to KLH and injected subcutaneously (into the flank;s.c.) or intradermally (i.d.) into wild-type mice (mice do not have CETPprotein), CETP-tg mice, or rabbits, respectively, and induced immuneresponse to the injected peptide with all adjuvants that were tested(Alum and CFA (Complete Freund's adjuvant); Gerbu).

For all in vitro inhibiting mimotopes listed above, antibodies reactingto the original CETP epitope could be detected in mice and in rabbits.

For 5 out of 6 mimotopes (see below and Table 1) antibodies reactingwith purified human CETP and recombinantly expressed human CETP could bedetected in ELISAs from rabbit sera:

F2-9 C SFLDTLT (SEQ ID NO. 45) F3-6 C NFLKTLS (SEQ ID NO. 46) F3-18C DFLRTLT (SEQ ID NO. 47) F3-34 C TFLSSLA (SEQ ID NO. 49) F3-38C GFLDSLM (SEQ ID NO. 50)

Subcutaneous injections in the flank were performed in week 1, week 3and week 7 with 30 μg peptide-KLH per mouse. Intradermal injections inthe ear were performed in week 1, week 3 and week 6 with 10 μgpeptide-KLH per mouse. Sera were taken 2 weeks after the 3rd injection.Vaccine formulation with Alum (always 1 mg per mouse): up to 250 μl,injected into one flank. The Alum formulation with 1 ml per mouse (500μl into each flank) was in 1×PBS as buffer.

Vaccine formulation with Gerbu Adjuvant 100 (Gerbu Cat. Nr. #3100;always 50 μl adjuvant per mouse): 200 μl, 100 μl injected into eachflank comprising 1×HEPES as buffer.

TABLE 1 Results of the titer determination P4073 injected (FGFPEH-p irrel- Adjuvant KLH mimotope LLVDFLOSLS) evant Alum s.c. (30 KLH1:20.000 n.a. 1:400 no titer μg peptide) p4073-KLH C- 1:70.000 n.a.1:20.000 no titer FGFPEHLLVD- FLQSLS (SEQ ID NO. 147) p4223-KLH F2-9; C-1:15.000 1:15.000 1:6.400 no titer SFLDTLT (SEQ ID NO. 45) p4181-KLHF3-6 C- 1:8.000 1:6.400 1:800 no titer NFLKTLS (SEQ ID NO. 46) p4184-KLHF3-18 C- 1:5.000 1:10.000 1:3.000 1:2.500 DFLRTLT (SEQ ID NO. 47) p4187F3-34 C- 1:3.200 1:9.000 1:4.000 no titer TFLSSLA (SEQ ID NO. 49)p4188-KLH F3-38 C- 1:10.000 1:9.000 1.5.000 no titer GFLDSLM (SEQID NO. 50) p4227-KLH P12-19; C- 1:12.800 1:10.000 1:5.000 no titerSANPRDFLETLF (SEQ ID NO. 55) p4228-KLH P12-21; C- 1:10.000 1:4.0001:1.000 1:400 RMFPESFLDTLW (SEQ ID NO. 56) KLH/Gerbu s.c. KLH 1:70.000n.a. 1:6.000 1:800 (30 μg pep- tide) p4073-KLH C- 1:25.000 n.a. 1:15.0001:200 FGFPEHLLVD- FLQSLS (SEQ ID NO. 147) p4223-KLH F2-9; C- 1:40.0001:25.000 1:50.000 1:1.000 SFLDTLT (SEQ ID NO. 45) p4181-KLH F3-6 C-1:20.000 1.20.000 1:8.000 1:400 NFLKTLS (SEQ ID NO. 46) p4184-KLHF3-18 C- 1:27.000 1.35.000 1:15.000 1:6.000 DFLRTLT (SEQ ID NO. 47)p4187-KLH F3-34 C- 1.20.000 1.20.000 1:15.000 no titer TFLSSLA (SEQID NO. 49) p4188-KLH F3-38 C- 1:40.000 1:35.000 1:35.000 1:400GFLDSLM (SEQ ID NO. 50) p4227-KLH P12-19; C- 1.20.000 1:30.000 1.3.0001:400 SANPRDFLETLF (SEQ ID NO. 55) p4228-KLH P12-21; C- 1:27.000 1:8.0001:5.000 no titer RMFPESFLDTLW (SEQ ID NO. 56) p4073-KLH C- 1:10.0001:10.000 no titer FGFPEHLLVD- FLQSLS (SEQ ID NO. 147) KLH/Alum i.d. KLH1:12.800 n.a. no titer no titer (10 μg pep- tide) p4073-KLH C- 1:10.000n.a. 1:3.200 no titer FGFPEHLLVD- FLQSLS (SEQ ID NO. 147) p4223-KLHF2-9; C- 1:6.400 1:3.200 SFLDTLT (SEQ ID NO. 45) p4181-KLH F3-6 C-1:10.000 1:1.500 1:600 no titer NFLKTLS (SEQ ID NO. 46) p4184-KLHF3-18 C- 1:15.000 1:5.000 1:1.500 no titer DFLRTLT (SEQ ID NO. 47)p4187-KLH F3-34 C- 1:50.000 1:6.400 1:3.200 1:500 TFLSSLA (SEQID NO. 49) p4188-KLH F3-38 C- 1:12.000 1:5.000 1:2.000 no titerGFLDSLM (SEQ ID NO. 50) p4227-KLH P12-19; C- 1:6.400 1:6.400 no titerno titer SANPRDFLETLF (SEQ ID NO. 55) p4228-KLH P12-21; C- 1:20.0001:2.000 1:1.600 no titer RMFPESFLDTLW (SEQ ID NO. 56) p4298-KLHFr12/3/84ext2; 1:25.000 1:3.200 1:1.600 no titer C- FGFPAHVSID-WLQSLS (SEQ ID NO. 184)

3.1.2. Phage Display Library Ph.D. 12

3.1.2.1. Screening with Monoclonal Antibody “Paula”

Out of 20 amino acid sequences derived from this screen, 3 wereinhibiting in in vitro inhibition experiments:

P12-19 SANPRDFLETLF (SEQ ID NO. 55) P12-21 RMFPESFLDTLW (SEQ ID NO. 56)P12-37 TIYDSFLDSLAS (SEQ ID NO. 57)

Not inhibiting peptides were:

P12-5/44/46/49 HQSDDKMPWWFF (SEQ ID NO. 27) P12-9 KPYLLKDFLEAL(SEQ ID NO. 58) P12-24/43-_ AMGPYDALDLFL (SEQ ID NO. 59) P12-25TWNPIESFLESL (SEQ ID NO. 60) P12-28 + 42 YVWQDPSFTTFF (SEQ ID NO. 28)P12-30 QYQTPLTFLEAL (SEQ ID NO. 61) P12-35- RHISPATFLEAL (SEQ ID NO. 62)P12-39- HTDSFLSTFYGD (SEQ ID NO. 63) P12-42- YVWQDPSFTTFF(SEQ ID NO. 29) P12-45- ADSTFTSFLQTL (SEQ ID NO. 64) P12-50-_GPVSIYADTDFL (SEQ ID NO. 65) P12-51-_ DSNDTLTLAAFL (SEQ ID NO. 66)P12-52-_ NGSPALSHMLFL (SEQ ID NO. 33) P12-53- TDYDPMWVFFGY(SEQ ID NO. 34) P12-56- IFPLDSQWQTFW (SEQ ID NO. 35) P12-58-NESMPDLFYQPS (SEQ ID NO. 36) P12-61- DWGDKYFSSFWN (SEQ ID NO. 37)

Results of 2 typical competition ELISAs are shown in FIGS. 2A and 2 b.

All 3 mimotopes were coupled to KLH and injected into wildtype mice(mice do not have CETP protein), CETP-tg mice, or rabbits, respectively,and induced immune response to the injected peptide with all adjuvantsthat were tested (Alum and CFA; Gerbu).

Mimotope P12-19; C-SANPRDFLETLF (SEQ ID NO. 55) and P12-21;C-RMFPESFLDTLW (SEQ ID NO. 56) induced an immune response to theoriginal CETP epitope in wt mice and in rabbits.

In contrast thereto, mimotope P12-37 C-TIYDSFLDSLAS (SEQ ID NO. 57) didnot induce an antibody response to the original epitope.

3.2 Screening with 2 Antibodies Derived from “Fusion I”: “Frida” and“James”

3.2.1. Phage Display Library pH.D. 7

3.2.1.1. Screening with Monoclonal Antibodies “Frida” and “James”

Two different peptide sequences were identified in these screens, 11 of12 clones that were sequenced had identical sequences. These peptidesare not inhibiting in in vitro competition experiments.

Fr7-2-2 Fr7-2B-65 Fr7-3-7 Fr7-3-13 Fr7-3-26 Fr7-3-32 Ja7-2-22 Ja7-3-28Ja7-3-41 Ja7-3-52 Ja7-3-56 VSAYNNV (SEQ ID NO. 38) Ja7-3-89 WPLHLWQ(SEQ ID NO. 39)

The results of 2 representative competition ELISAs with mAb “Frida” areshown in FIGS. 3A and 3 b. The same pattern was seen with mAb “James”.

3.2.2. Phage Display Library pH.D. 12

3.2.2.1. Screening with Monoclonal Antibody “Frida”

Fr12/2/6 TPTHYYADFSQL (SEQ ID NO. 67) Fr12/2/11 LPGHLIWDSLHY(SEQ ID NO. 68) Fr12/2/27 LPQTHPLHLLED (SEQ ID NO. 69) Fr12/3/1Fr12/3/19 Fr12/3/88 IPYHHLVDQLHH (SEQ ID NO. 70) Fr12/3/26 Fr12/3/65YPYHVQVDVLQN (SEQ ID NO. 71) Fr12/3/68 IPSHHLQDSLQL (SEQ ID NO. 72)Fr12/3/12 EYAHHTSLDLRQ (SEQ ID NO. 73) Fr12/3/83 EPLHFRSDRIQA(SEQ ID NO. 74) Fr12/3/55 ATPSHLIIDRAQ (SEQ ID NO. 75) Fr12/3/63APKHLYADMSQA (SEQ ID NO. 76) Fr12/3/84 FKPAHVSIDWLQ (SEQ ID NO. 77)Fr12/3/47 MPAHLSRDLRQS (SEQ ID NO. 78) Fr12/3/80 NPKHYSIDRHQA(SEQ ID NO. 79) Fr12/3/40 SPQHLTTDRAQA (SEQ ID NO. 80) Fr12/3/35TPFHFAQDSWQW (SEQ ID NO. 81)

None of the 15 amino acid sequences identified in this screen wereinhibiting in in vitro competition experiments. However, sequenceanalysis revealed rather high homology to the original protein sequencefor many of the mimotopes. On the other hand, for some peptides bindingof monoclonal antibody “Frida” to ELISA plates coated with mimotope-BSAcould be shown (see FIGS. 4 a and 4 b).

This shows that binding of monoclonal antibody to immobilised mimotopesdoes not necessarily allow to predict inhibition in in vitro competitionELISA.

In vitro inhibition experiments with variations of the original sequenceFGFPEHLLVDFLQSLS (SEQ ID NO. 147) (16 C-terminal AA of CETP protein)showed that removing more than 2 amino acids from the N-terminus or morethan 1 amino acid from the C-terminus abolishes inhibition (formonoclonal antibodies “Frida” and “James”. “Paula” and “Felix” recognisea different part of the original sequence).

In addition, simultaneously removing 2 amino acids from the N-terminusand 1 amino acid from the C-terminus also results in a peptide that isnot inhibiting in vitro any more.

(SEQ ID NO. 147) C-FGFPEHLLVDFLQSLS “original” sequence (peptide derivedfrom CETP)/inhibiting in vitro C-GFPEHLLVDFLQSLS sequence N-1/inhibiting in vitro C-FPEHLLVDFLQSLS sequence N-2/ inhibiting in vitroC-PEHLLVDFLQSLS sequence N-3/ evtl. slightly inhibiting in vitroC-FGFPEHLLVDFLQSL sequence C-1/ inhibiting in vitroC-FGFPEHLLVDFLQS sequence C-2/ not inhibiting in vitroC-FPEHLLVDFLQSL sequence N-2 and C-1/not inhibiting in vitro! “original”FGFPEHLLVDFLQSLS Fr12/2/6 TPTHYYADFSQL (SEQ ID NO. 67) Fr12/2/11LPGHLIWDSLHY (SEQ ID NO. 68) Fr12/2/27 LPQTHPLHLLED (SEQ ID NO. 69)Fr12/3/1 IPYHHLVDQLHH (SEQ ID NO. 70) Fr12/3/19 IPYHHLVDQLHH(SEQ ID NO. 70) Fr12/3/88 IPYHHLVDQLHH (SEQ ID NO. 70) Fr12/3/26YPYHVQVDVLQN (SEQ ID NO. 71) Fr12/3/65 YPYHVQVDVLQN (SEQ ID NO. 71)Fr12/3/68 IPSHHLQDSLQL (SEQ ID NO. 72) Fr12/3/12 EYAHHTSLDLRQ(SEQ ID NO. 73) Fr12/3/83 EPLHFRSDRIQA (SEQ ID NO. 74) Fr12/3/55ATPSHLIIDRAQ (SEQ ID NO. 75) Fr12/3/63 APKHLYADMSQA (SEQ ID NO. 76)Fr12/3/84 FKPAHVSIDWLQ (SEQ ID NO. 77) Fr12/3/47 MPAHLSRDLRQS(SEQ ID NO. 78) Fr12/3/80 NPKHYSIDRHQA (SEQ ID NO. 79) Fr12/3/40SPQHLTTDRAQA (SEQ ID NO. 80) Fr12/3/35 TPFHFAQDSWQW (SEQ ID NO. 81)

Consequently, using the original CETP sequence as a template, peptidesequences obtained in this Phage Display procedure were elongated on theN-terminus and/or C-terminus to check whether in vitro inhibition ispossible with longer peptides.

3.2.2.2. Mimotopes Frida pH.D.12 and Variations Thereof:

Fr12/2/6 TPTHYYADFSQL (SEQ ID NO. 67) Fr12/2/6 ext1 TPTHYYADFSQLLS(SEQ ID NO. 82) Fr12/2/6 ext2 TPTHYYADFSQSLS (SEQ ID NO. 83)Fr12/2/6 ext3 GTPTHYYADFSQLL (SEQ ID NO. 84) Fr12/2/6 ext4GTPTHYYADFSQSL (SEQ ID NO. 85) Fr12/2/6 ext5 FGTPTHYYADFSQSLS(SEQ ID NO. 86) Fr12/2/6 ext6 FGFPTHYYADFSQSLS (SEQ ID NO. 87) Fr12/2/11LPGHLIWDSLHY (SEQ ID NO. 68) Fr12/2/11 ext1 LPGHLIWDSLHYL(SEQ ID NO. 89) Fr12/2/11 ext2 LPGHLIWDSLHYLS (SEQ ID NO. 90)Fr12/2/11 ext3 LPGHLIWDSLHSL (SEQ ID NO. 91) Fr12/2/11 ext4LPGHLIWDSLHSLS (SEQ ID NO. 92) Fr12/2/11 ext5 GLPGHLIWDSLHYL(SEQ ID NO. 93) Fr12/2/11 ext5 GLPGHLIWDSLHSL (SEQ ID NO. 94)Fr12/2/11 ext6 FGLPGHLIWDSLHSLS (SEQ ID NO. 95) Fr12/2/11 ext7FGFPGHLIWDSLHSLS (SEQ ID NO. 96) Fr12/2/27 LPQTHPLHLLED (SEQ ID NO. 69)Fr12/3/1/19/88  IPYHHLVDQLHLS (SEQ ID NO. 99) ext1 Fr12/3/1/19/88 IPYHHLVDQLHSLS (SEQ ID NO. 100) ext2 Fr12/3/1/19/88  FGIPYHHLVDQLHHLS(SEQ ID NO. 101) ext3 Fr12/3/1/19/88  FGFPYHHLVDQLHSLS (SEQ ID NO. 102)ext4 Fr12/3/26/65ext1 YPYHVQVDVLQNLS (SEQ ID NO. 104) Fr12/3/26/65ext2YPYHVQVDVLQSLS (SEQ ID NO. 105) Fr12/3/26/65ext3 FGYPYHVQVDVLQNLS(SEQ ID NO. 106) Fr12/3/26/65ext4 FGFPYHVQVDVLQSLS (SEQ ID NO. 107)Fr12/3/68 ext1 IPSHHLQDSLQLLS (SEQ ID NO. 109) Fr12/3/68 ext2IPSHHLQDSLQSLS (SEQ ID NO. 110) Fr12/3/68 ext3 GIPSHHLQDSLQLL(SEQ ID NO. 111) Fr12/3/68 ext4 FGIPSHHLQDSLQLLS (SEQ ID NO. 112)Fr12/3/68 ext5 FGFPSHHLQDSLQSLS (SEQ ID NO. 113) Fr12/3/83 ext1EPLHFRSDRIQALS (SEQ ID NO. 116) Fr12/3/83 ext2 EPLHFRSDRIQSLS (SEQ ID NO. 117) Fr12/3/83 ext3 GEPLHFRSDRIQAL (SEQ ID NO. 118)Fr12/3/83 ext4 FGEPLHFRSDRIQALS (SEQ ID NO. 119) Fr12/3/83 ext5FGFPLHFRSDRIQSLS (SEQ ID NO. 120) Fr12/3/55 ext1 ATPSHLIIDRAQSLS(SEQ ID NO. 176) Fr12/3/55 ext2 FGFPSHLIIDRAQSLS (SEQ ID NO. 177)Fr12/3/55 ext2  FGFPSHLIIDWAQSLS (SEQ ID NO. 178) R->W Fr12/3/55 ext2 FGFPSHLIIDWLQSLS (SEQ ID NO. 179) RA->WL Fr12/3/63 ext1 APKHLYADMSQALS(SEQ ID NO. 122) Fr12/3/63 ext2 APKHLYADMSQSLS (SEQ ID NO. 123)Fr12/3/63 ext3 GAPKHLYADMSQAL (SEQ ID NO. 124) Fr12/3/63 ext4FGFPKHLYADMSQSLS (SEQ ID NO. 125) Fr12/3/84 ext1 FKPAHVSIDWLQSLS(SEQ ID NO. 183) Fr12/3/84 ext2 FGFPAHVSIDWLQSLS (SEQ ID NO. 184)Fr12/3/47 ext1 MPAHLSRDLRQSL (SEQ ID NO. 127) Fr12/3/47 ext2MPAHLSRDLRQSLS (SEQ ID NO. 128) Fr12/3/47 ext3 GMPAHLSRDLRQSL(SEQ ID NO. 129) Fr12/3/47 ext4 FGFPAHLSRDLRQSLS (SEQ ID NO. 130)Fr12/3/40 ext1 SPQHLTTDRAQALS (SEQ ID NO. 168) Fr12/3/40 ext2SPQHLTTDRAQSLS (SEQ ID NO. 169) Fr12/3/40 ext3 GSPQHLTTDRAQAL(SEQ ID NO. 170) Fr12/3/40 ext4 FGFPQHLTTDRAQSLS (SEQ ID NO. 171)Fr12/3/35 ext1 TPFHFAQDSWQWLS (SEQ ID NO. 133) Fr12/3/35 ext2TPFHFAQDSWQSLS (SEQ ID NO. 134) Fr12/3/35 ext3 GTPFHFAQDSWQWL(SEQ ID NO. 135) Fr12/3/35 ext4 FGFPFHFAQDSWQSLS (SEQ ID NO. 136)

Representative examples of inhibition ELISA are shown in FIGS. 5A and 5b. The elongated peptides Fr12/3/84 ext2 and Fr12/3/55 ext3 showed asignificant inhibition:

C-FGFPSHLIIDRAQSLS Fr12/3/55 ext3 (SEQ ID NO. 177) C-FGFPAHVSIDWLQSLSFr12/3/84 ext2 (SEQ ID NO. 184)

Three additional peptides were also inhibiting in this assay:

C-FGFPYHVQVDVLQSLS Fr12/3/26/65ext4 (SEQ ID NO. 107) C-FKPAHVSIDWLQSLSFr12/3/84 ext1 (SEQ ID NO. 183) C-FGFPQHLTTDRAQSLS Fr12/3/40 ext4(SEQ ID NO. 171)

After sequence analysis comparing the original epitope and all mimotopesderived from Phage Display screens additional 2 peptides were created.

For mimotope Fr12/3/55 ext3 C-FGFPSHLIIDRAQSLS (SEQ ID NO. 177)(inhibiting in ELISA, see above) amino acid exchanges were tested ininhibition ELISA:

Strongly inhibiting: C-FGFPAHVSIDWLQSLS (SEQ ID NO. 184) Fr12/3/84 ext2Slightly inhibiting: C-FGFPSHLIIDRAQSLS (SEQ ID NO. 177) Fr12/3/55 ext3Peptides with altered  sequences (inhibiting, see FIG. 6):C-FGFPSHLIIDWAQSLS (SEQ ID NO. 178) Fr12/3/55 ext2 W instead of RC-FGFPSHLIIDWLQSLS (SEQ ID NO. 179) Fr12/3/55 ext2 WL instead  of RAFurther preferred mimotopes have been characterised by the followingexample-set-up:

Exp. Nr. C42-1 KLH/Alum — CETP-42 C42-2 p4073-KLH/ C-FGFPEHLLVDFLQSLSAlum (SEQ ID NO. 147 C42-3 p4073 LLV->LFV p4468-KLH/ C-FGFPEHLFVDFLQSLSAlum (SEQ ID NO. 252) C42-4 Fr12/3/84 ext2 VSI->VFI P4361-KLH/C-FGFPAHVFIDWLQSLS Alum (SEQ ID NO. 222) C42-5 Fr12/3/84 ext2 VSI->VHIp4469- C-FGFPAHVHIDWLQSLS KLH/Alum (SEQ ID NO. 253) C42-6Fr12/3/84 ext2 VSI->VPI p4470- C-FGFPAHVPIDWLQSLS KLH/Alum(SEQ ID NO. 254) C42-7 Fr12/3/84 ext2 VSI->VWI p4471- C-FGFPAHVWIDWLQSLSKLH/Alum (SEQ ID NO. 229) C42-8 Fr12/3/55 ext2 R->W LII->LFI p4472-C-FGFPSHLFIDWAQSLS KLH/Alum (SEQ ID NO. 255) C42-9Fr12/3/84 ext2 VSöVFI FGF- p4473- C-PGFPAHVFIDWLQLIT >PGF SLS->LITKLH/Alum (SEQ ID NO. 256) C42-10 Fr12/3/84 ext2 VSI->VYI P4362-KLH/C-FGFPAHVYIDWLQSLS Alum (SEQ ID NO. 223) Exp. Nr. C45-1 KLH/Alum —CETP-45 C45-2 p1358-KLH/ neg. control peptide Alum C45-3 p4073-KLH/C-FGFPEHLLVDFLQSLS Alum (SEQ ID NO. 147) C45-4Fr12/3/84 ext2 VSI->VFI SLS- p4474-KLH/ C-FGFPAHVFIDWLQSLN >SLN Alum(SEQ ID NO. 230) C45-5 Fr12/3/84 ext2 VSI->FSI p4475-KLH/C-FGFPAHFSIDWLQSLS Alum (SEQ ID NO. 231) C45-6 Fr12/3/84 ext2 VSI->VSFp4476-KLH/ C-FGFPAHVSFDWLQSLS Alum (SEQ ID NO. 232) C45-7Fr12/3/84 ext2 VSI->VFI PAH- p4477-KLH/ C-FGFPEHVFIDWLQSLS >PEH Alum(SEQ ID NO. 233) C45-8 Fr12/3/1/19/88 ext4 p4284-KLH/ C-FGFPYHHLVDQLHSLSAlum (SEQ ID NO. 102) C45-9 Fr12/3/84 ext1 VSI->VFI plus  p4479-KLH/C-GFKPAHVFIDWLQSLS G on N-terminus Alum (SEQ ID NO. 270) C45-10Fr12/3/84 ext2 VSI->VFI plus  p4480-KLH/ C-DFGFPAHVFIDWLQSLSD on N-terminus; =4361 plus D Alum (SEQ ID NO. 234) C45-11Fr12/3/40 ext4 RA->WL LTT->LFT p4481-KLH/ C-FGFPQHLFTDWLQSLS=p4369 with exchange TöF Alum (SEQ ID NO. 237) C45-12Fr12/3/55 ext2 RA->WL (see C- p4325-KLH/ C-FGFPSHLIIDWLQSLS31 and C-33; sera inhibiting Alum (SEQ ID NO. 179) activity) C45-13Fr12/3/84 ext2 FGF->FYF (see p4343-KLH/ C-FYFPAHVSIDWLQSLSC-33: recogn. protein/not in- Alum (SEQ ID NO. 204) hibiting activity)C45-14 rabbit sequence p4125-KLH/ C-FGFPKHLLVDFLQSLS Alum(SEQ ID NO. 238)

3.2.2.3. In Vivo Testing of Mimotopes

Female Balb/c mice, five mice per group, were subcutaneously immunizedwith 30 μg peptide coupled to KLH. Control groups were administered KLHor C-FGFPEHLLVDFLQSLS (SEQ ID NO. 147). As adjuvant alum was used. Thepeptides administered were all able to bind to “Frida” and to induce animmune response for CETP, although some of these peptides did notinhibit the binding of CETP to “Frida” in vitro (in an in vitroinhibition assay). The in vitro ELISA assay to determine the antibodytiter was performed with pooled sera after two vaccinations in a twoweek interval (S2; see FIGS. 7 a to 7 d). The wells of the ELISA platewere coated with KLH (positive control), mimotope-BSA conjugate,C-FGFPEHLLVDFLQSLS (SEQ ID NO. 147) and a irrelevant peptide-BSAconjugate (negative control). The detection was performed withanti-mouse IgG.

3.2.3. Phage Display Library pH.D. 7C7

3.2.3.1. Screening with Monoclonal Antibodies “Frida” and “James”

Fr2-1 ACSFAYLYRC (SEQ ID NO. 137) Fr2-5 Fr2-6 Fr2-18 Fr2-19 Fr2-28 Ja2-5Ja2-20 Ja2-23 Ja2-24 Ja2-30 ACFMGDKWVC (SEQ ID NO. 138) Fr2-7 Fr2-9ACVLYPKAIC (SEQ ID NO. 139) Fr2-11 Ja2-19 ACYMGQQFVC (SEQ ID NO. 140)Fr2-16 ACLTAYLHWC (SEQ ID NO. 141) Fr2-20 ACTLFPVAYC (SEQ ID NO. 142)Fr2-25 ACWLFPYAHC (SEQ ID NO. 143) Fr2-26 ACKSINMWLC (SEQ ID NO. 144)Fr2-27 ACQTINRWLC (SEQ ID NO. 145)

Due to their cyclic nature of these mimotope-peptides their synthesis ismore complicated than the synthesis of linear peptides. Seven out of 9cyclic sequences were chosen for in vitro analysis in inhibition ELISA(see FIGS. 8 a and 8 b). None of these sequences inhibited binding ofthe monoclonal antibody that was used for Phage Display Screening to theoriginal CETP epitope. In addition, when these peptides were coupled toBSA and coated onto ELISA plate they were not detected by the monoclonalantibody (see FIG. 9). This was in contrast to data with mimotopesderived from the Ph.D.7 or Ph.D.12 libraries, where the monoclonalantibodies bound to most of the identified mimotopes when these peptideswere coupled to BSA and coated onto ELISA plates.

Example 3 CETP Activity Assay

The CETP activity assay was performed with assays commercially available(e.g. ROAR CETP Activity Assay) and described, for instance, in the U.S.Pat. No. 5,585,235, U.S. Pat. No. 5,618,683 and U.S. Pat. No. 5,770,355.The assay is performed according to the manufacturers' recommendations.

1. Peptide consisting of at least one amino acid sequence selected fromthe group consisting of SYHATFL, TMAFPLN, HYHGAFL, EHHDIFL, SSLELFL,TGLSVFL, WMPSLFY, SMPWWFF, TMPLLFW, DTWPGLE, SMPPIFY, MPLWWWD, SMPNLFY,RMPPIFY, NPFEVFL, TLPNWFW, SMPLTFY, SFLDTLT, NFLKTLS, DFLRTLT, AFLDTLV,TFLSSLA, GFLDSLM, SPHPHFL, NFMSIGL, SQFLASL, SNFLKTL, TGFLATL, WSWPGLN,IAWPGLD, SKFMDTL, SDFLRAL, SMPMVFY, YEWVGLM, KGFLDHL, SANPRDFLETLF,RMFPESFLDTLW, TIYDSFLDSLAS, HQSDDKMPWWFF, KPYLLKDFLEAL, AMGPYDALDLFL,TWNPIESFLESL, YVWQDPSFTTFF, QYQTPLTFLEAL, RHISPATFLEAL, HTDSFLSTFYGD,YVWQDPSFTTFF, ADSTFTSFLQTL, GPVSIYADTDFL, DSNDTLTLAAFL, NGSPALSHMLFL,TDYDPMWVFFGY, IFPLDSQWQTFW, NESMPDLFYQPS, DWGDKYFSSFWN, VSAYNNV,WPLHLWQ, TPTHYYADFSQL, LPGHLIWDSLHY, LPQTHPLHLLED, IPYHHLVDQLHH,YPYHVQVDVLQN, IPSHHLQDSLQL, EYAHHTSLDLRQ, EPLHFRSDRIQA, ATPSHLIIDRAQ,APKHLYADMSQA, FKPAHVSIDWLQ, MPAHLSRDLRQS, NPKHYSIDRHQA, SPQHLTTDRAQA,TPFHFAQDSWQW, TPTHYYADFSQLLS, TPTHYYADFSQSLS, GTPTHYYADFSQLL,GTPTHYYADFSQSL, FGTPTHYYADFSQSLS, FGFPTHYYADFSQSLS, LPGHLIWDSLHY,LPGHLIWDSLHYL, LPGHLIWDSLHYLS, LPGHLIWDSLHSL, LPGHLIWDSLHSLS,GLPGHLIWDSLHYL, GLPGHLIWDSLHSL, FGLPGHLIWDSLHSLS, FGFPGHLIWDSLHSLS,LPQTHPLHLLED, IPYHHLVDQLHH, IPYHHLVDQLHLS, IPYHHLVDQLHSLS,FGIPYHHLVDQLHHLS, FGFPYHHLVDQLHSLS, YPYHVQVDVLQN, YPYHVQVDVLQNLS,YPYHVQVDVLQSLS, FGYPYHVQVDVLQNLS, FGFPYHVQVDVLQSLS, IPSHHLQDSLQL,IPSHHLQDSLQLLS, IPSHHLQDSLQSLS, GIPSHHLQDSLQLL, FGIPSHHLQDSLQLLS,FGFPSHHLQDSLQSLS, EYAHHTSLDLRQ, EPLHFRSDRIQA, EPLHFRSDRIQALS,EPLHFRSDRIQSLS, GEPLHFRSDRIQAL, FGEPLHFRSDRIQALS, FGFPLHFRSDRIQSLS,APKHLYADMSQA, APKHLYADMSQALS, APKHLYADMSQSLS, GAPKHLYADMSQAL,FGFPKHLYADMSQSLS, MPAHLSRDLRQS, MPAHLSRDLRQSL, MPAHLSRDLRQSLS,GMPAHLSRDLRQSL, FGFPAHLSRDLRQSLS, NPKHYSIDRHQA, TPFHFAQDSWQW,TPFHFAQDSWQWLS, TPFHFAQDSWQSLS, GTPFHFAQDSWQWL, FGFPFHFAQDSWQSLS,ACSFAYLYRC, ACFMGDKWVC, ACVLYPKAIC, ACYMGQQFVC, ACLTAYLHWC, ACTLFPVAYC,ACWLFPYAHC, ACKSINMWLC, ACQTINRWLC, FGFPEHLLVDFLQSLS, FGFPEHLLVDFLQSLS,FPEHLLVDFLQSL, AGFPEHLLVDFLQSLS, FAFPEHLLVDFLQSLS, FGAPEHLLVDFLQSLS,FGFAEHLLVDFLQSLS, FGFPAHLLVDFLQSLS, FGFPEALLVDFLQSLS, FGFPEHALVDFLQSLS,FGFPEHLAVDFLQSLS, FGFPEHLLADFLQSLS, FGFPEHLLVAFLQSLS, FGFPEHLLVDALQSLS,FGFPEHLLVDFAQSLS, FGFPEHLLVDFLASLS, FGFPEHLLVDFLQALS, FGFPEHLLVDFLQSAS,FGFPEHLLVDFLQSLA, FAFPAHLLVDFLQALA, AAFPAHLLADFLQALA, SPQHLTTDRAQA,SPQHLTTDRAQALS, SPQHLTTDRAQALS, GSPQHLTTDRAQAL, FGFPQHLTTDRAQSLS,FGFPQHLTTDWAQSLS, FGFPQHLTTDRLQSLS, FGFPQHLTTDWLQSLS, ATPSHLIIDRAQ,ATPSHLIIDRAQSLS, FGFPSHLIIDRAQSLS, FGFPSHLIIDWAQSLS, FGFPSHLIIDWLQSLS,FGFPSHLIIDWSQSLS, FATPSHLIIDWLQSLS, FKPAHVSIDWLQ, FKPAHVSIDWLQSLS,FGFPAHVSIDWLQSLS, AGFPAHVSIDWLQSLS, FAFPAHVSIDWLQSLS, FGAPAHVSIDWLQSLS,FGFAAHVSIDWLQSLS, FGFPAHVSADWLQSLS, FGFPAHVSIDWLQALS, FGFPAHVSIDWLQSLA,FAFPAHVSIDWLQALA, FGFAAHVSIDWLQSLS, FGFFAHVSIDWLQSLS, FGFPAHVSIRWLQSLS,FGFPAHVSIEWLQSLS, FGFPAHVSIDWLNSLS, FGFPAHVSIDWLHSLS, AGFPAHVSIDWLQSLS,PGFPAHVSIDWLQSLS, WGFPAHVSIDWLQSLS, FAFPAHVSIDWLQSLS, FGFPAHVSIDWLQSLS,FYFPAHVSIDWLQSLS, FDFPAHVSIDWLQSLS, FGAPAHVSIDWLQSLS, FGFPAHVSIDWLQLLS,FGFPAHVSIDWLQWLS, FGFPAHVSIDWLQNLS, FGFPAHVSIDWLQTLS, FGFPAHVSIDWLQYLS,FGFPAHVSIDWLQSIS, FGFPAHVSIDWLQSLT, FGFPAHVSIDWLQSLY, FAFPAHVSIDWLQALA,FGFPAHVSIDRAQSLS, FGFPTHVSIDWLQSLS, FGFPFHVSIDWLQSLS, FGFPAHISIDWLQSLS,FGFPAHIIIDWLQSLS, FGFPAHLTTDWLQSLS, FGFPAHVFIDWLQSLS, FGFPAHVYIDWLQSLS,FGFPAHVSLDWLQSLS, FGFPAHVSADWLQSLS, TPTHYYADFSQSLS, FGFPAHVSIDWSQSLS,FGFPAHVSIDFSQSLS, FGFPSHIIIDWLQSLS, FGFPSHLIIEWLQSLS, AAFPAHLLADAAQALA,AAFPAHAAADFLQALA, AAFAAHLLADFLQAAA, AAAPAHLLVDAAQAAA, FAFPAHVFIDWLQSLS;FGFPAHVFIDWLQALS, FGFPAHVFIDWLQSLA, GFPAHVFIDWLQSLS, FPAHVFIDWLQSLS,PAHVFIDWLQSLS, FAFPAHVFIDWLQALA, FGFPEHLFVDFLQSLS, FGFPAHVHIDWLQSLS,FGFPAHVPIDWLQSLS, FGFPSHLFIDWAQSLS, PGFPAHVFIDWLQLIT, PAHVYIDWLQSLS,FGFPAHVYIDWLQ, FGFPAHVFIDWLQ, DFGFPSHLIIDWLQSLS, DFGFPAHVFIDWLQSLN,PSHLIIDWLQ, PAHVFIDWLQ, DFGFPAHVTIDWLQSLN, DFGFPAHVLIDWLQSLN,FGFPAHVYIDWLQSLS, FGFPAHVFIDWLQSLN and FGFPAHVFIDWLQSLA. 2.Pharmaceutical formulation comprising at least one peptide according toclaim
 1. 3. Formulation according to claim 2, characterised in that thepeptide is coupled to a pharmaceutically acceptable carrier, preferablyKLH (Keyhole Limpet Hemocyanin).